Polysubstituted benzimidazoles as antiviral agents

ABSTRACT

This invention relates to novel polysubstituted benzimidazoles and compositions and their use in the treatment of viral infections. The polysubstituted benzimidazoles and compositions of the present invention exhibit antiviral properties against viruses of the herpes family, particularly human cytomegalovirus (HCMV) and herpes simplex viruses (HSV).

SPONSORSHIP

This invention was made with government support under Contract No. NO1Al 42554 and NO1 Al 72641 awarded by the National Institute of Allergyand Infectious Diseases of the National Institutes of Health. Thegovernment has certain rights in this invention.

RELATED APPLICATIONS

This application is a divisional application of U.S. patent applicationSer. No. 08/050,470, filed May 3, 1993, now issued U.S. Pat. No.5,574,058; which is a continuation-in-part application of U.S. patentapplication Ser. No. 07/607,899, filed Nov. 1, 1990, now issued U.S.Pat. No. 5,248,672.

FIELD OF THE INVENTION

The present invention relates generally to polysubstitutedbenzimidazolas and, more particularly, to novel polysubstitutedbenzimidazoles and compositions, their preparation and their use asantiviral agents, particularly against human cytomegalovirus and herpessimplex virus.

BACKGROUND OF THE INVENTION

Antiviral activity of polysubstituted benzimidazolas such as5,6-dichloro-1-(β-D-ribofuranosyl) benzimidazole (DRB) and some closelyrelate derivatives has been previously described. Their activity againstspecific viruses, such as RNA rhinovirus and DNA herpes simplex virustype 1 and type 2, has also been reported.

Benzimidazole nucleosides are particularly attractive as potentialantiviral agents because of their stability toward some major pathwaysof bioactive purine (bicyclic) nucleoside inactivation, e.g.,deamination by adenosine deaminase and glycosidic bond cleavage bypurine nucleoside phosphorylases. Benzimidazole nucleosides such as DRBhave, however, demonstrated only marginal levels of activity orgenerally unacceptable levels of cytotoxicity, or both, thereby greatlydiminishing their usefulness in the treatment of viral infections. Itwould thus be desirable to provide polysubstituted benzimidazoles andcompositions thereof having good antiviral properties, preferably with alow degree of cytotoxicity.

SUMMARY OF THE INVENTION

The present Invention relates to novel antiviral compositions comprisinga polysubstituted benzimidazole and a pharmaceutically acceptablecarrier and methods of treatment therewith, wherein the polysubstitutedbenzimidazole is selected from the group consisting of compounds havingthe following formula and pharmaceutically acceptable salts andformulations thereof: ##STR1## wherein R₁ is H, R₂ is Cl, R₃ is Cl, R₄is H, R₅ is Cl and R₆ is β-D-ribofuranosyl (denoted compound 45 in thetext);

R₁ is H, R₂ is Cl, R₃ is Cl, R₄ is H, R₅ is Br and R₆ isβ-D-ribofuranosyl (denoted compound 52 in the text);

R₁ is H, R₂ is NO₂, R₃ is NO₂, R₄ is H, R₅ is Cl and R₆ isβ-D-ribofuranosyl (denoted compound 61 in the text);

R₁ is Cl, R₂ is H, R₃ is Cl, R₄ is H, R₅ is Cl and R₆ isβ-D-ribofuranosyl (denoted compound 81 in the text);

R₁ is H, R₂ is Cl, R₃ is Cl, R₄ is H, R₅ is I and R₆ isβ-D-ribofuranosyl (denoted compound 83a in the text);

R₁ is Br, R₂ is Br, R₃ is H, R₄ is H, R₅ is Cl and R₆ isβ-D-ribofuranosyl (denoted compound 85 in the text);

R₁ is H, R₂ is Br, R₃ is Cl, R₄ is H, R₅ is Cl and R₆ isβ-D-ribofuranosyl (denoted compound 95 in the text);

R₁ is H, R₂ is Cl, R₃ is Br, R₄ is H, R₅ is Cl and R₆ isβ-D-ribofuranosyl (denoted compound 99 in the text);

R₁ is H, R₂ is I, R₃ is I, R₄ is H, R₅ is Cl and R₆ is β-D-ribofuranosyl(denoted compound 107 in the text);

R₁ is H, R₂ is Cl, R₃ is Cl, R₄ is H, R₅ is Cl and R₆ is2'-deoxy-β-D-erythro-pentofuranosyl (denoted compound 111 in the text);

R₁ is H, R₂ is Cl, R₃ is Cl, R₄ is H, R₅ is Br and R₆ is2'-deoxy-β-D-erythro-pentofuranosyl (denoted compound 112 in the text);

R₁ is H, R₂ is Cl, R₃ is Cl, R₄ is H, R₅ is Br and R₆ is H (denotedcompound 7 in the text);

R₁ is H, R₂ is Cl, R₃ is F, R₄ is H, R₅ is Cl and R₆ is H (denotedcompound 12c in the text);

R₁ is Cl, R₂ is Cl, R₃ is Cl, R₄ is H, R₅ is Cl and R₆ is H (denotedcompound 13 in the text);

R₁ is H, R₂ is NO₂, R₃ is H, R₄ is H, R₅ is Cl and R₆ is H (denotedcompound 19 in the text);

R₁ is H, R₂ is I, R₃ is NO₂, R₄ is H, R₅ is Cl and R₆ is H (denotedcompound 26 in the text);

R₁ is Cl, R₂ is H, R₃ is Cl, R₄ is H, R₅ is Cl and R₆ is H (denotedcompound 32 in the text);

R₁ is H, R₂ is I, R₃ is I, R₄ is H, R₅ is Cl and R₆ is H (denotedcompound 41 in the text);

R₁ is Cl, R₂ is H, R₃ is CF₃, R₄ is H, R₅ is Cl and R₆ is H (denotedcompound 41c in the text);

R₁ is H, R₂ is Cl, R₃ is Cl, R₄ is H, R₅ is NH₂ and R₆ isβ-D-ribofuranosyl (denoted compound 44 in the text);

R₁ is H, R₂ is Cl, R₃ is Cl, R₄ is H, R₅ is SCH₂ C₆ H₅ and R₆ isβ-D-ribofuranosyl (denoted compound 54 in the text);

R₁ is H, R₂ is Br, R₃ is Br, R₄ is H, R₅ is Cl and R₆ isβ-D-ribofuranosyl (denoted compound 57 in the text);

R₁ is H, R₂ is F, R₃ is F, R₄ is H, R₅ is Cl and R₆ is β-D-ribofuranosyl(denoted compound 65 in the text);

R₁ is H, R₂ is Cl, R₃ is F, R₄ is H, R₅ is Cl and R₆ isβ-D-ribofuranosyl (denoted compound 65a in the text);

R₁ is H, R₂ is H, R₃ is Cl, R₄ is H, R₅ is Cl and R₆ isβ-D-ribofuranosyl (denoted compound 67 in the text);

R₁ is Cl, R₂ is H, R₃ is Cl, R₄ is H, R₅ is CF₃ and R₆ isβ-D-ribofuranosyl (denoted compound 81b in the text);

R₁ is Cl, R₂ is H, R₃ is CF₃, R₄ is H, R₅ is Cl and R₆ isβ-D-ribofuranosyl (denoted compound 81c in the text);

R₁ is H, R₂ is Br, R₃ is H, R₄ is H, R₅ is Cl and R₆ isβ-D-ribofuranosyl (denoted compound 87 in the text);

R₁ is H, R₂ is H, R₃ is Br, R₄ is H, R₅ is Cl and R₆ isβ-D-ribofuranosyl (denoted compound 90 in the text);

R₁ is Cl, R₂ is Cl, R₃ is Cl, R₄ is H, R₅ is Cl and R₆ isβ-D-ribofuranosyl (denoted compound 92 in the text);

R₁ is Br, R₂ is Br, R₃ Br, R₄ is H, R₅ is Cl and R₆ is β-D-ribofuranosyl(denoted compound 103 in the text);

R₁ is H, R₂ is Cl, R₃ is Cl, R₄ is H, R₅ is NH₂ and R₆ is2'-deoxy-β-D-erythro-pentofuranosyl (denoted compound 113 in the text);

R₁ is H, R₂ is Cl, R₃ is Cl, R₄ is H, R₅ is Cl and R₆ isβ-D-arabinofuranosyl (denoted compound 134 in the text);

R₁ is Cl, R₂ is Cl, R₃ is Cl, R₄ is Cl, R₅ is Cl and R₆ is(1,3-dihydroxy-2-propoxy)methyl (denoted compound 155 in the text);

R₁ is Cl, R₂ is Cl, R₃ is Cl, R₄ is Cl, R₅ is NH₂ and R₆ is (1,3-dihydroxy-2-propoxy)methyl (denoted compound 156 in the text);

R₁ is Cl, R₂ is Cl, R₃ is Cl, R₄ is Cl, R₅ is Cl and R₆ is2-hydroxyethoxymethyl (denoted compound 166 in the text);

R₁ is Cl, R₂ is Cl, R₃ is Cl, R₄ is Cl, R₅ is OCH₃ and R₆ is2-hydroxyethoxymethyl (denoted compound 166a in the text);

R₁ is Cl, R₂ is Cl, R₃ is Cl, R₄ is Cl, R₅ is NH₂ and R₆ is2-hydroxyethoxymethyl (denoted compound 167 in the text);

R₁ is H, R₂ is Cl, R₃ is Cl, R₄ is H, R₅ is NH₂ and R₆ is benzyl(denoted compound 182 in the text);

R₁ is H, R₂ is Cl, R₃ is Cl, R₄ is H, R₅ is Cl and R₆ is5-O-acetyl-β-D-ribofuranosyl (denoted compound 42a in the text);

R₁ is H, R₂ is Cl, R₃ is Cl, R₄ is H, R₅ is Br and R₆ is5-O-acetyl-β-D-ribofuranosyl (denoted compound 52b in the text);

R₁ is H, R₂ is Cl, R₃ is Cl, R₄ is H, R₅ is Cl and R₆ is2,3,5-tri-O-acetyl-β-D-ribofuranosyl (denoted compound 42 in the text);

R₁ is H, R₂ is Cl, R₃ is Cl, R₄ is H, R₅ is Br and R₆ is2,3,5-tri-O-acetyl-β-D-ribofuranosyl (denoted compound 52a in the text);

and operative combinations thereof.

As used herein, by "pharmaceutically acceptable carrier" is meant anycomposition, solvent, dispersion medium, coating, delivery vehicle orthe like, which can be employed to administer the compounds andcompositions of the present invention without undue adversephysiological effects. By "operative combination" is meant anychemically compatible combination of the compounds which does noteliminate the antiviral activity of the composition.

The present invention also relates to a method of antiviral treatmentgenerally comprising the step of administering to the viral host atherapeutically effective amount of a polysubstituted benzimidazoleselected from the above-described group. By "therapeutically effectiveamount" is meant an amount generally effective to achieve the desiredantiviral effect.

The present invention further relates to the use of a polysubstitutedbenzimidazole selected from the above-described group in themanufacturing of medicaments for antiviral use. The present inventionalso relates to compositions comprising a polysubstituted benzimidazoleselected from the above-described group in combination with otherantiviral agents outside the group.

The present invention further relates to the novel polysubstitutedbenzimidazoles used in the compositions and treatments of the presentinvention and methods of their making. Novel benzimidazoles includecompounds 52, 61, 83a, 85, 95, 99, 107, 111, 112, 7, 12c, 13, 19, 26,32, 41, 41c, 57, 65, 65a, 67, 81b, 81c, 87, 90, 92, 103, 113, 134, 182,42a, 52b, 42, 52a, 81, 155, 156, 166, 166a, 167.

BRIEF DESCRIPTION OF THE DRAWINGS AND TABLES

FIGS. 1, 1a, 1b, 2, 2a, 2b, 2c, 2d, and 2e are synthesis schemes andsubstituent charts of polysubstituted benzimidazoles in accordance withand illustrating the principles of the present invention.

FIG. 3 is a dose response curve comparing the activity against humancytomegalovirus of two polysubstituted benzimidazole nucleosides(compounds 45 and 111) to the known drug ganciciovir in accordance withthe principles of the present invention.

FIG. 4 is a graph illustrating the low degree of cytotoxicity (cellgrowth inhibition) of a polysubstituted benzimidazole nucleoside(compound 45) in accordance with the principles of the presentinvention.

FIG. 5 is a graph illustrating the reversibility of the cytotoxiceffects produced by a very high level of a polysubstituted benzimidazolenucleoside (compound 45) in accordance with the principles of thepresent invention.

FIG. 6 is a graph illustrating the cytotoxic effects of DRB on cellgrowth and its irreversibility.

Table 1 illustrates the antiviral activity and cytotoxicity ofpolysubstituted benzimidazoles in accordance with the principles of thepresent invention.

Table 2 illustrates the effects of a polysubstituted benzimidazolenucleoside on the replication of selected herpes viruses in accordancewith the principles of the present invention.

Table 3 illustrates the antiviral activity and cytotoxicity ofpolysubstituted benzimidazoles in accordance with the principles of thepresent invention.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

A. Chemical Structure of Polysubstituted Benzimidazoles of the Invention

Novel antiviral compositions of the present invention comprise apolysubstituted benzimidazole and a pharmaceutically acceptable carrier,wherein the polysubstituted benzimidazole is selected from the groupconsisting of compounds having the following formula andpharmaceutically acceptable salts and formulations thereof: ##STR2##wherein R₁ is H, R₂ is Cl, R₃ is Cl, R₄ is H, R₅ is Cl and R₆ isβ-D-ribofuranosyl (denoted compound 45 in the text);

R₁ is H, R₂ is Cl, R₃ is Cl, R₄ is H, R₅ is Br and R₆ isβ-D-ribofuranosyl (denoted compound 52 in the text);

R₁ is H, R₂ is NO₂, R₃ is NO₂, R₄ is H, R₅ is Cl and R₆ isβ-D-ribofuranosyl (denoted compound 61 in the text);

R₁ is Cl, R₂ is H, R₃ is Cl, R₄ is H, R₅ is Cl and R₆ isβ-D-ribofuranosyl (denoted compound 81 in the text);

R₁ is H, R₂ is Cl, R₃ is Cl, R₄ is H, R₅ is I and R₆ isβ-D-ribofuranosyl (denoted compound 83a in the text);

R₁ is Br, R₂ is Br, R₃ is H, R₄ is H, R₅ is Cl and R₆ isβ-D-ribofuranosyl (denoted compound 85 in the text);

R₁ is H, R₂ is Br, R₃ is Cl, R₄ is H, R₅ is Cl and R₆ isβ-D-ribofuranosyl (denoted compound 95 in the text);

R₁ is H, R₂ is Cl, R₃ is Br, R₄ is H, R₅ is Cl and R₆ isβ-D-ribofuranosyl (denoted compound 99 in the text);

R₁ is H, R₂ is I, R₃ is I, R₄ is H, R₅ is Cl and R₆ is β-D-ribofuranosyl(denoted compound 107 in the text);

R₁ is H, R₂ is Cl, R₃ is Cl, R₄ is H, R₅ is Cl and R₆ is2'-deoxy-β-D-erythro-pentofuranosyl (denoted compound 111 in the text);

R₁ is H, R₂ is Cl, R₃ is Cl, R₄ is H, R₅ is Br and R₆ is2'-deoxy-β-D-erythro-pentofuranosyl (denoted compound 112 in the text);

R₁ is H, R₂ is Cl, R₃ is Cl, R₄ is H, R₅ is Br and R₆ is H (denotedcompound 7 in the text);

R₁ is H, R₂ is Cl, R₃ is F, R₄ is H, R₅ is Cl and R₆ is H (denotedcompound 12c in the text);

R₁ is Cl, R₂ is Cl, R₃ is Cl, R₄ is H, R₅ is Cl and R₆ is H (denotedcompound 13 in the text);

R₁ is H, R₂ is NO₂, R₃ is H, R₄ is H, R₅ is Cl and R₆ is H (denotedcompound 19 in the text);

R₁ is H, R₂ is I, R₃ is NO₂, R₄ is H, R₅ is Cl and R₆ is H (denotedcompound 26 in the text);

R₁ is Cl, R₂ is H, R₃ is Cl, R₄ is H, R₅ is Cl and R₆ is H (denotedcompound 32 in the text);

R₁ is H, R₂ is I, R₃ is I, R₄ is H, R₅ is Cl and R₆ is H (denotedcompound 41 in the text);

R₁ is Cl, R₂ is H, R₃ is CF₃, R₄ is H, R₅ is Cl and R₆ is H (denotedcompound 41c in the text);

R₁ is H, R₂ is Cl, R₃ is Cl, R₄ is H, R₅ is NH₂ and R₆ isβ-D-ribofuranosyl (denoted compound 44 in the text);

R₁ is H, R₂ is Cl, R₃ is Cl, R₄ is H, R₅ is SCH₂ C₆ H₅ and R₆ isβ-D-ribofuranosyl (denoted compound 54 in the text);

R₁ is H, R₂ is Br, R₃ is Br, R₄ is H, R₅ is Cl and R₆ isβ-D-ribofuranosyl (denoted compound 57 in the text);

R₁ is H, R₂ is F, R₃ is F, R₄ is H, R₅ is Cl and R₆ is β-D-ribofuranosyl(denoted compound 65in the text);

R₁ is H, R₂ is Cl, R₃ is F, R₄ is H, R₅ is Cl and R₆ isβ-D-ribofuranosyl (denoted compound 65a in the text);

R₁ is H, R₂ is H, R₃ is Cl, R₄ is H, R₅ is Cl and R₆ isβ-D-ribofuranosyl (denoted compound 67 in the text);

R₁ is Cl, R₂ is H, .R₃ is Cl, R₄ is H, R₅ is CF₃ and R₆ isβ-D-ribofuranosyl (denoted compound 81b in the text);

R₁ is Cl, R₂ is H, R₃ is CF₃, R₄ is H, R₅ is Cl and R₆ isβ-D-ribofuranosyl (denoted compound 81c in the text);

R₁ is H, R₂ is Br, R₃ is H, R₄ is H, R₅ is Cl and R₆ isβ-D-ribofuranosyl (denoted compound 87 in the text);

R₁ is H, R₂ is H, R₃ is Br, R₄ is H, R₅ is Cl and R₆ isβ-D-ribofuranosyl (denoted compound 90 in the text);

R₁ is Cl, R₂ is Cl, R₃ is Cl, R₄ is H, R₅ is Cl and R₆ isβ-D-ribofuranosyl (denoted compound 92 in the text);

R₁ is Br, R₂ is Br, R₃ is Br, R₄ is H, R₅ is Cl and R₆ isβ-D-ribofuranosyl (denoted compound 103 in the text);

R₁ is H, R₂ is Cl, R₃ is Cl, R₄ is H, R₅ is NH₂ and R₆ is2'-deoxy-β-D-erythro-pentofuranosyl (denoted compound 113 in the text);

R₁ is H, R₂ is Cl, R₃ is Cl, R₄ is H, R₅ is Cl and R₆ isβ-D-arabinofuranosyl (denoted compound 134 in the text);

R₁ is Cl, R₂ is Cl, R₃ is Cl, R₄ is Cl, R₅ is Cl and R₆ is(1,3-dihydroxy-2-propoxy)methyl (denoted compound 155 in the text);

R₁ is Cl, R₂ is Cl, R₃ is Cl, R₄ is Cl, R₅ is NH₂ and R₆ is(1,3-dihydroxy-2-propoxy)methyl (denoted compound 156 in the text);

R₁ is Cl, R₂ is Cl, R₃ is Cl, R₄ is Cl, R₅ is Cl and R₆ is2-hydroxyethoxymethyl (denoted compound 166 in the text);

R₁ is Cl, R₂ is Cl, R₃ is Cl, R₄ is Cl, R₅ is OCH₃ and R₆ is2-hydroxyethoxymethyl (denoted compound 166a in the text);

R₁ is Cl, R₂ is Cl, R₃ is Cl, R₄ is Cl, R₅ is NH₂ and R₆ is2-hydroxyethoxymethyl (denoted compound 167 in the text);

R₁ is H, R₂ is Cl, R₃ is Cl, R₄ is H, R₅ is NH₂ and R₆ is benzyl(denoted compound 182 in the text);

R₁ is H, R₂ is Cl, R₃ is Cl, R₄ is H, R₅ is Cl and R₆ is5-O-acetyl-β-D-ribofuranosyl (denoted compound 42a in the text);

R₁ is H, R₂ is Cl, R₃ is Cl, R₄ is H, R₅ is Br and R₆ is5-O-acetyl-β-D-ribofuranosyl (denoted compound 52b in the text);

R₁ is H, R₂ is Cl, R₃ is Cl, R₄ is H, R₅ is Cl and R₆ is2,3,5-tri-O-acetyl-β-D-ribofuranosyl (denoted compound 42 in the text);

R₁ is H, R₂ is Cl, R₃ is Cl, R₄ is H, R₅ is Br and R₆ is2,3,5-tri-O-acetyl-β-D-ribofuranosyl (denoted compound 52a in the text);

and operative combinations thereof.

Thus compounds in the practice of the compositions and methods of thepresent invention include:

2,5,6-trichloro-1-(β-D-ribofuranosyl)benzimidazole where R₁, R₄ =H; R₂,R₃, R₅ =Cl; R₆ =β-D-ribofuranosyl (denoted compound 45 in the text);

5,6-dichloro-2-bromo-1-(β-D-ribofuranosyl)benzimidazole where R₁, R₄ =H;R₂, R₃ =Cl; R₅ =Br; R₆ =β-D-ribofuranosyl (denoted compound 52 in thetext);

2-chloro-5,6-dinitro-1-(β-D-ribofuranosyl)benzimidazole where R₁, R₄ =H;R₂, R₃ =NO₂ ; R₅ =Cl; R₆ =β-D-ribofuranosyl (denoted compound 61 in thetext);

2,4,6-trichloro-1 -(β-D-ribofuranosyl)benzimidazole where R₂, R₄ =H; R₁,R₃, R₅ =Cl; R₆ =β-D-ribofuranosyl (denoted compound 81 in the text);

5,6-dichloro-2-iodo-1-(β-D-ribofuranosyl)benzimidazole where R₁, R₄ =H;R₂, R₃ =Cl; R₅ =I; R₆ =β-D-ribofuranosyl (denoted compound 83a in thetext);

2-chloro-4,5-dibromo-1 -(β-D-ribofuranosyl)benzimidazole where R₃, R₄=H; R₁, R₂ =Br; R₅ =Cl; R₆ =β-D-ribofuranosyl (denoted compound 85 inthe text);

5-bromo-2,6-dichloro-1-(β-D-ribofuranosyl)benzimidazole where R₁, R₄ =H;R₂ =Br; R₃, R₅ =Cl; R₆ =β-D-ribofuranosyl (denoted compound 95 in thetext);

6-bromo-2,5-dichloro-1-(β-D-ribofuranosyl)benzimidazole where R₁, R₄ =H;R₂, R₅ =Cl; R₃ =Br; R₆ =β-D-ribofuranosyl (denoted compound 99 in thetext);

2-chloro-5,6-diiodo-1-(β-D-ribofuranosyl)benzimidazole where R₁, R₄ =H;R₂, R₃ =I; R₅ =Cl; R₆ =β-D-ribofuranosyl (denoted compound 107 in thetext);

2,5,6-trichloro-1-(2'-deoxy-β-D-erythro-pentofuranosyl)benzimidazolewhere R₁, R₄ =H; R₂, R₃, R₅ =Cl; R₆ =2'-deoxy-β-D-erythro-pentofuranosyl(denoted compound 111 in the text);

5,6-dichloro-2-bromo-1-(2'-deoxy-β-D-erythro-pentofuranosyl)benzimidazolewhere R₁, R₄ =H; R₂, R₃ =Cl; R₅ =Br; R₆=2'-deoxy-β-D-erythro-pentofuranosyl (denoted compound 112 in the text);

2-bromo-5,6-dichiorobenzimidazole where R₁, R₄, R₆ =H; R₂, R₃ =Cl; R₅=Br (denoted compound 7 in the text);

2,5(6)-dichloro-6(5)-fluorobenzimidazole where R₁, R₄, R₆ =H; R₂, R₅=Cl; R₃ =F (denoted compound 12c in the text);

2,4,5,6-tetrachlorobenzimidazole where R₄, R₆ =H; R₁, R₂, R₃, R₅ =Cl(denoted compound 13 in the text);

2-chloro-5-nitrobenzimidazole where R₁, R₃, R₄, R₆ =H; R₂ =NO₂ ; R₅ =Cl(denoted compound 19 in the text);

2-chloro-5(6)-iodo-6(5)-nitrobenzimidazole where R₁, R₄, R₆ =H; R₂ =I;R₃ =NO₂ ; R₅ =Cl (denoted compound 26 in the text);

2,4,6-trichlorobenzimidazole where R₂, R₄, R₆ =H; R₁, R₃, R₅ =Cl(denoted compound 32 in the text);

2-chloro-5,6-diiodobenzimidizole where R₁, R₄, R₆ =H; R₂, R₃ =I, R₅ =Cl(denoted compound 41 in the text);

2,4(7)-dichloro-6(5)-trifluoromethylbenzimidazole where R₁, R₄, R₆ =H;R₂, R₃ =I; R₅ =Cl (denoted compound 41c in the text);

2-amino-5,6-dichloro-1-(β-D-ribofuranosyl)benzimidazole where R₁, R₄ =H;R₂, R₃ =Cl; R₅ =NH₂ ; R₆ =(β-D-ribofuranosyl (denoted compound 44 in thetext); 2-benzylthio-5,6-dichloro-1-(β-D-ribofuranosyl)benzimidazolewhere R₁, R₄ =H; R₂, R₃ =Cl; R₅ =SCH₂ C₆ H₅ ; R₆ =β-D-ribofuranosyl(denoted compound 54 in the text);2-chloro-5,6-dibromo-1-(β-D-ribofuranosyl)benzimidazole where R₁,R₄ =H;R₂, R₃

=Br; R₅ =Cl; R₆ =β-D-ribofuranosyl (denoted compound 57 in the text);2-chloro-5,6-difluoro-1-(β-D-ribofuranosyl)benzimidazole where R₁, R₄=H; R₂, R₃

=F; R₅ =Cl; R₆ =β-D-ribofuranosyl (denoted compound 65 in the text);2,5-dichloro-6-fluoro-1-β-D-ribofuranosylbenzimidazole (2,5) and2,6-dichloro-5-fluoro-1-β-D-ribofuranosylbenzimidazole (2,6) where R₁,R₄ =H; R₂, R₅ =Cl; R₃ =F; R₆ =β-D-ribofuranosyl (denoted compound 65a inthe text);

2,6-dichloro-1-(β-D-ribofuranosyl)benzimidazole where R₁, R₂, R₄ =H; R₃,R₅ =Cl; R₆ =β-D-ribofuranosyl (denoted compound 67 in the text);

4,6-dichloro-2-trifluoromethyl-1-(β-D-ribofuranosyl)benzimidazole whereR₂, R₄ =H; R₁, R₃ =Cl; R₅ =CF₃ ; R₆ =β-D-ribofuranosyl (denoted compound81b in the text);

2,4-dichloro-1(β-D-ribofuranosyl)-6-trifluoromethylbenzimidazole whereR₂, R₄ =H; R₁, R₅ =Cl; R₃ =CF₃ ; R₆ =β-D-ribofuranosyl (denoted compound81c in the text);

5-bromo-2-chloro-1-(β-D-ribofuranosyl)benzimidazole where R₁, R₃, R₄ =H;R₂ =Br; R₅ =Cl; R₆ =β-D-ribofuranosyl (denoted compound 87 in the text);

6-bromo-2-chloro-1-(β-D-ribofuranosyl)benzimidazole where R₁, R₂, R₄ =H;R₃ =Br; R₅ =Cl, R₆ =β-D-ribofuranosyl (denoted compound 90 in the text);

1-(β-D-ribofuranosyl)-2,4,5,6-tetrachlorobenzimidazole where R₄ =H; R₁,R₂, R₃, R₅ =Cl; R₆ =β-D-ribofuranosyl (denoted compound 92 in the text);

2-chloro-1-(β-D-ribofuranosyl)-4,5,6-tribromobenzimidazole where R₄ =H;R₁, R₂, R₃ =Br; R₅ =Cl; R₆ =β-D-ribofuranosyl (denoted compound 103 inthe text);

2-amino-5,6-dichloro-1-(2-deoxy-β-D-erythro-pentofuranosyl)benzimidazolewhere R₁, R₄ =H; R₂, R₃ =Cl; R₅ =NH₂ ; R₆ =2'-deoxy-β-D-pentofuranosyl(denoted compound 113 in the text);

2,5,6-trichloro-1-(β-D-arabinofuranosyl)benzimidazole where R₁, R₄ =H;R₂, R₃, R₅ =Cl; R₆ =β-D-arabinofuranosyl (denoted compound 134 in thetext);

2,4,5,6,7-pentachloro-1- (1,3-dihydroxy-2-propoxy)methyl!benzimidazotewhere R₁, R₂, R₃, R₄, R₅ =Cl; R₆ =(1,3-dihydroxy-2-propoxy)methyl(denoted compound 155 in the text);

2-amino-4,5,6,7-tetrachloro-1-(1,3-dihydroxy-2-propoxy)methyl!benzimidazole where R₁, R₂, R₃, R₄ =Cl;R₅ =NH₂ ; R₆ =(1,3-dihydroxy-2-propoxy)methyl (denoted compound 156 inthe text);

2,4,5,6,7-pentachloro-1-(2-hydroxyethoxymethyl)benzimidazole where R₁,R₂, R₃, R₄, R₅ =Cl; R₆ =2-hydroxyethoxymethyl) (denoted compound 166 inthe text);

2-methoxy-4,5,6,7-tetrachloro-1-(2-hydroxyethoxymethyl)benzimidazolewhere R₁, R₂, R₃, R₄ =Cl; R₅ =OCH₃ ; R₆ =2-hydroxyethoxymethyl (denotedcompound 166a in the text);

2-amino-4,5,6,7-tetrachloro-1-(2-hydroxyethoxymethyl)benzimidazole whereR₁, R₂, R₃, R₄ =Cl; R₅ =NH₂ ; R₆ =2-hydroxyethoxymethyl (denotedcompound 167 in the text);

2-amino-1-benzyl-5,6-dichlorobenzimidazole where R₁, R₄ =H; R₂, R₃ =Cl;R₅ =NH₂ ; R₆ =benzyl (denoted compound 182 in the text);

1-(5-O-acetyl-β-D-ribofuranosyl)-2,5,6-trichlorobenzimidazole where R₁,R₄ =H; R₂, R₃, R₅ =Cl; R₆ =5-O-acetyl-β-D-ribofuranosyl (denotedcompound 42a in the text);

1-(5-O-acetyl-β-D-ribofuranosyl)-2-bromo-5,6-dichlorobenzimidazole whereR₁, R₄ =H; R₂, R₃ =Cl; R₅ =Br; R₆ =5-O-acetyl-β-D-ribofuranosyl (denotedcompound 52b in the text);

2,5,6-trichioro-1-(2,3,5-tri-O-acetyl-β-D-ribofuranosyl)benzimidazolewhere R₁, R₄ =H; R₂, R₃, R₅ =Cl; R₆=2,3,5-tri-O-acetyl-(β-D-ribofuranosyl (denoted compound 42 in thetext);

2-bromo-5,6-dichloro-1-(2,3,5-tri-O-acetyl-β-D-ribofuranosyl)benzimidazolewhere R₁, R₄ =H; R₂, R₃ =Cl; R₅ =Br; R₆=2,3,5-tri-O-acetyl-β-D-ribofuranosyl (denoted compound 52a in thetext);

and operative combinations thereof.

Preferred compounds in the practice of the compositions and methods ofthe present invention include:

2,5,6-trichloro-1-(β-D-ribofuranosyl)benzimidazole where R₁, R₄ =H; R₂,R₃, R₅ =Cl; R₆ =β-D-ribofuranosyl (denoted compound 45 in the text);

5,6-dichloro-2-bromo-1-(β-D-ribofuranosyl)benzimidazole where R₁, R₄ =H;R₂, R₃ =Cl; R₅ =Br; R₆ =β-D-ribofuranosyl (denoted compound 52 in thetext);

2-chloro-5,6-dinitro-1-(β-D-ribofuranosyl)benzimidazole where R₁, R₄ =H;R₂, R₃ =NO₂ ; R₅ =Cl; R₆ =β-D-ribofuranosyl (denoted compound 61 in thetext);

2,4,6-trichloro-1-(β-D-ribofuranosyl)benzimidazole where R2, R₄ =H; R₁,R₃, R₅ =Cl; R₆ =β-D-ribofuranosyl (denoted compound 81 in the text);

5,6-dichloro-2-iodo-1-(β-D-ribofuranosyl)benzimidazole where R₁, R₄ =H;R₂, R₃ =Cl; R₅ =I; R₆ =β-D-ribofuranosyl (denoted compound 83a in thetext);

2-chloro-4,5-dibromo-1-(β-D-ribofuranosyl)benzimidazole where R₃, R₄ =H;R₁, R₂ =Br; R₅ =Cl; R₆ =β-D-ribofuranosyl (denoted compound 85 in thetext);

5-bromo-2,6-dichloro-1-(β-D-ribofuranosyl)benzimidazole where R₁, R₄ =H;R₂ =Br; R₃, R₅ =Cl; R₆ =β-D-ribofuranosyl (denoted compound 95 in thetext);

6-bromo-2,5-dichloro-1-(β-D-ribofuranosyl)benzimidazole where R₁, R₄ =H;R₂, R₅ =Cl; R₃ =Br; R₆ =β-D-ribofuranosyl (denoted compound 99 in thetext);

2-chloro-5,6-diiodo-1-(β-D-ribofuranosyl)benzimidazole where R₁, R₄ =H;R₂, R₃ =I; R₅ =Cl; R₆ =β-D-ribofuranosyl (denoted compound 107 in thetext);

2,5,6-trichloro-1-(2'-deoxy-β-D-erythro-pentofuranosyl)benzimidazolewhere R₁, R₄ =H; R₂, R₃, R₅ =Cl; R₆ =2'-deoxy-β-D-erythro-pentofuranosyl(denoted compound 111 in the text);

5,6-dichloro-2-bromo-1-(2'-deoxy-β-D-erythro-pentofuranosyl)benzimidazolewhere R₁, R₄ =H; R₂, R₃ =Cl; R₅ =Br; R₆=2'-deoxy-β-D-erythro-pentofuranosyl (denoted compound 112 in the text);

2-bromo-5,6-dichlorobenzimidazole where R₁, R₄, R₆ =H; R₂, R₃ =Cl; R₅=Br (denoted compound 7 in the text);

2-chloro-5-nitrobenzimidazole where R₁, R₃, R₄, R₆ =H; R₂ =NO₂ ; R₅ =Cl(denoted compound 19 in the text);

2-chloro-5(6)-iodo-6(5)-nitrobenzimidazole where R₁, R₄, R₆ =H; R₂ =I;R₃ =NO₂ ; R₅ =Cl (denoted compound 26 in the text);

2,4,6-trichloroloenzimidazole where R₂, R₄, R₆ =H; R₁, R₃, R₅ =Cl(denoted compound 32 in the text);

2,benzylthio-5,6-dichloro-1-(β-D-ribofuranosyl)benzimidazole where R₁,R₄ =H; R₂, R₃ =Cl; R₅ =SCH₂ C₆ H₅ ; R₆ =β-D-ribofuranosyl (denotedcompound 54 in the text);

2-chloro-5,6-dibromo-1-(β-D-ribofuranosyl)benzimidazote where R₁, R₄ =H;R₂, R₃ =Br; R₅ =Cl; R₆ =β-D-ribofuranosyl (denoted compound 57 in thetext);

2-chloro-5,6-difluoro-1-(β-D-ribofuranosyl)benzimidazole where R₁, R₄=H; R₂, R₃ =F; R₅ =Cl; R₆ =β-D-ribofuranosyl (denoted compound 65 in thetext);

2,5-dichloro-6-fluoro-1-β-D-ribofuranosylbenzimidazole (2,5) and2,6-dichloro-5-fluoro-1-β-D-ribofuranosylbenzimidazole (2,6) where R₁,R₄ =H; R₂, R₅ =Cl; R₃ =F; R₆ =β-D-ribofuranosyl (denoted compound 65a inthe text);

2,6-dichloro-1-(β-D-ribofuranosyl)benzimidazole where R₁, R₂, R₄ =H; R₃,R₅ =Cl; R₆ =β-D-ribofuranosyl (denoted compound 67 in the text);

4,6-dichloro-2-trifluoromethyl-1-(β-D-ribofuranosyl)benzimidazole whereR₂, R₄ =H; R₁, R₃ =Cl; R₅ =CF₃ ; R₆ =β-D-ribofuranosyl (denoted compound81b in the text);

2,4-dichloro-1-(β-D-ribofuranosyl)-6-trifluoromethylbenzimidazole whereR₂, R₄ =H; R₁, R₅ =Cl; R₃ =CF₃ ; R₆ =β-D-ribofuranosyl (denoted compound81c in the text);

5-bromo-2-chloro-1-(β-D-ribofuranosyl)benzimidazole where R₁, R₃, R₄ =H;R₂ =Br; R₅ =Cl; R₆ =β-D-ribofuranosyl (denoted compound 87 in the text);

6-bromo-2-chloro-1-(β-D-ribofuranosyl)benzimidazole where R₁, R₂, R₄ =H;R₃ =Br; R₅ =Cl, R₆ =β-D-ribofuranosyl (denoted compound 90 in the text);

1-(β-D-ribofuranosyl)-2,4,5,6-tetrachlorobenzimidazole where R₄ =H; R₁,R₂, R₃, R₅ =Cl; R₆ =β-D-ribofuranosyl (denoted compound 92 in the text);

2-chloro-1-(β-D-ribofuranosyl)-4,5,6-tribromobenzimidazole where R₄ =H;R₁, R₂, R₃ =Br; R₅ =Cl; R₆ =β-D-ribofuranosyl (denoted compound 103 inthe text);

2-amino-5,6-dichloro-1-(2-deoxy-β-D-erythro-pentofuranosyl)benzimidazolewhere R₁, R₄ =H; R₂, R₃ =Cl; R₅ =NH₂ ; R₆=2'-deoxy-β-D-erythro-pentofuranosyl (denoted compound 113 in the text);

2,5,6-trichloro-1-(β-D-arabinofuranosyl)benzimidazole where R₁, R₄ =H;R₂, R₃, R₅ =Cl; R₆ =β-D-arabinofuranosyl (denoted compound 134 in thetext);

2,4,5,6,7-pentachloro-1- (1,3-dihydroxy-2-propoxy)methyl!benzimidazolewhere R₁, R₂, R₃, R₄, R₅ =Cl; R₆ =(1,3-dihydroxy-2-propoxy)methyl(denoted compound 155 in the text);

2-amino-4,5,6,7-tetrachloro-1-(1,3-dihydroxy-2-propoxy)methyl!benzimidazole where R₁, R₂, R₃, R₄ =Cl;R₅ =NH₂ ; R₆ =(1 ,3-dihydroxy-2-propoxy)methyl (denoted compound 156 inthe text);

2-methoxy-4,5,6,7-tetrachloro-1-(2-hydroxyethoxymethyl)benzimidazolewhere R₁, R₂, R₃, R₄ =Cl; R₅ =OCH₃ ; R₆ =2-hydroxyethoxymethyl (denotedcompound 166a in the text);

2-amino-4,5,6,7-tetrachloro-1-(2-hydroxyethoxymethyl)benzimidazole whereR₁, R₂, R₃, R₄ =Cl; R₅ =NH₂ ; R₆ =2-hydroxyethoxymethyl (denotedcompound 167 in the text);

2-amino-1-benzyl-5,6-dichlorobenzimidazole where R₁, R₄ =H; R₂ ; R₃ =Cl;R₅ =NH₂ ; R₆ =benzyl (denoted compound 182 in the text);

1-(5-O-acetyl-β-D-ribofuranosyl)-2,5,6-trichlorobenzimidazole where R₁,R₄ =H; R₂, R₃, R₅ =Cl; R₆ =5-O-acetyl-β-D-ribofuranosyl (denotedcompound 42a in the text);

1-(5-O-acetyl-β-D-ribofuranosyl)-2-bromo-5,6-dichlorobenzimidazole whereR₁, R₄ =H; R₂, R₃ =Cl; R₅ =Br; R₆ =5-O-acetyl-D-ribofuranosyl (denotedcompound 52b in the text);

2,5,6-trichloro-1-(2,3,5-tri-O-acetyl-β-D-ribofuranosyl)benzimidazolewhere R₁, R₄ =H; R₂, R₃, R₅ =Cl; R₆=2,3,5-tri-O-acetyl-β-D-ribofuranosyl (denoted compound 42 in the text);

2-bromo-5,6-dichloro-1-(2,3,5-tri-O-acetyl-β-D-ribofuranosyl)benzimidazolewhere R₁, R₄ =H; R₂, R₃ =Cl; R₅ =Br; R₆=2,3,5-tri-O-acetyl-β-D-ribofuranosyl (denoted compound 52a in thetext); and operative combinations thereof.

More preferred compounds in the practice of the compositions and methodsof the present invention include:

2,5,6-trichloro-1-(β-D-ribofuranosyl)benzimidazole where R₁, R₄ =H; R₂,R₃, R₅ =Cl; R₆ =β-D-ribofuranosyl (denoted compound 45 in the text);

5,6-dichloro-2-bromo-1-(β-D-ribofuranosyl)benzimidazole where R₁, R₄ =H;R₂, R₃ =Cl; R₅ =Br; R₆ =β-D-ribofuranosyl (denoted compound 52 in thetext);

2-chloro-4,5dibromo-1-(β-D-ribofuranosyl)benzimidazole where R₃, R₄ =H;R₁, R₂ =Br; R₅ =Cl; R₆ =β-D-ribofuranosyl (denoted compound 85 in thetext);

2,5,6-trichloro-1-(2'-deoxy-β-D-erythro-pentafuranosyl)benzimidazolewhere R₁, R₄ =H; R₂, R₃, R₅ =Cl; R₆ =2'-deoxy-β-D-erythro-pentofuranosyl(denoted compound 111 in the text);

2-benzylthio-5,6-dichloro-1-(=-D-ribofuranosyl)benzimidazole where R₁,R₄ =H; R₂, R₃ =Cl; R₅ =SCH₂ C₆ H₅ ; R₆ =β-D-ribofuranosyl (denotedcompound 54 in the text);

2-chloro-5,6-difluoro-1-(β-D-ribofuranosyl)benzimidazole where R₁, R₄=H; R₂, R₃ =F; R₅ =Cl; R₆ =β-D-ribofuranosyl (denoted compound 65 in thetext);

2,5-dichloro-6-fluoro-1-β-D-ribofuranosylbenzimidazole (2,5) and2,6-dichloro-5-fluoro-1-β-D-ribofuranosylbenzimidazole (2,6) where R₁,R₄ =H; R₂, R₅ =Cl; R₃ =F; R₆ =β-D-ribofuranosyl (denoted compound 65a inthe text);

2,4-dichloro-1-(β-D-ribofuranosyl)-6-trifluoromethylbenzimidazole whereR₂, R₄ =H; R₁, R₅ =Cl; R₃ =CF₃ ; R₆ =β-D-ribofuranosyl (denoted compound81c in the text);

5-bromo-2-chloro-1-(β-D-ribofuranosyl)benzimidazole where R₁, R₃, R₄ =H;R₂ =Br; R₅ =Cl; R₆ =β-D-ribofuranosyl (denoted compound 87 in the text);

6-bromo-2-chloro-1-(β-D-ribofuranosyl)benzimidazole where R₁, R₂, R₄ =H;R₃ =Br; R₅ =Cl; R₆ =β-D-ribofuranosyl (denoted compound 90 in the text);

1-(β-D-ribofuranosyl)-2,4,5,-tetrachlorobenzimidazole where R₄ =H; R₁,P₂, R₃, R₅ =Cl; R₆ =β-D-ribofuranosyl (denoted compound 92 in the text);

2-amino-4,5,6,7-tetrachloro-1-(1,3-dihydroxy-2-propoxy)methyl!benzimidazole where R₁, R₂, R₃, R₄ =Cl;R₅ =NH₂ ; R₆ =(1,3-dihydroxy-2-propoxy)methyl (denoted compound 156 inthe text);

2-amino-1-benzyl-5,6-dichlorobenzimidazole where R₁, R₄ =H; R₂, R₃ =Cl;R₅ =NH₂ ; R₆ =benzyl (denoted compound 182 in the text);

and operative combinations thereof.

The present invention also comprises novel polysubstitutedbenzimidazoles, and, as later described, their synthesis and use inmedicaments and methods of treatment, Novel polysubstitutedbenzimidazoles of the present invention include compounds 52, 61, 83a,85, 95, 99, 107, 111, 112, 7, 12c, 13, 19, 26, 32, 41, 41c, 57, 65, 65a,67, 81b, 81c, 87, 90, 92, 103, 113, 134, 182, 42a, 52b, 42, 52a, 81,155, 156, 166, 166a, 167 of the above-described group. The structures ofcompounds 81, 155, 166, 166, 166a, and 167 were depicted in Abstractssubmitted to the April 1990 meeting of the American Chemical Society inBoston, Mass. The structures of compounds 45, 44 and 54 were alsopreviously proposed in Townsend et al., Chem. Reviews 70:389 (1970) andSmith et al., Cancer Treat. Rep. 60:1567-1584 (1976), but without adescription of synthesis and without recognition of any antiviralactivity or therapeutic value as antiviral agents. Preferred novelcompounds of the present invention include compounds 52, 85, 111, 65,65a, 81c, 87, 90, 92 and 182.

B. Pharmaceutical Compositions and Administration

The polysubstituted benzimidazoles used in the practice of the presentinvention all exhibit antiviral activity, many with acceptablecytotoxicity. It will be appreciated that compounds of the presentinvention which exhibit relatively high antiviral activity versuscytotoxicity, i.e. good selectivity, are preferred. It will also beappreciated that antiviral treatment in accordance with the presentinvention encompasses the treatment of viral infections, as well asprophylactic treatment which may be desired in certain situations, e.g.in immuno-compromised patients, such as bone marrow transplant patients.

The polysubstituted benzimidazoles of the invention have beendemonstrated as particularly effective against viruses of the herpesfamily. They are thus useful in treatment against human cytomegalovirus(HCMV) and herpes simplex viruses types 1 and 2. Other virusescontemplated to be treated within the scope of the present inventioninclude, but are not limited to: varicella-zoster virus (varicella;zoster, chickenpox; shingles); Epstein-Barr virus (infectiousmononucleosis; glandular fever; and Burkittis lymphoma); HHV6; humanimmunodeficiency virus (HIV) and hepatitis viruses.

The compounds and compositions of the present invention can be used inthe manufacture of medicaments and in antiviral treatment of humans andother animals by administration in accordance with conventionalprocedures, such as an active ingredient in pharmaceutical compositions.The compounds of the invention can be provided as pharmaceuticallyacceptable formulations and/or "prodrugs," including but not limited toesters, especially carboxylic acid esters (preferably C₁ to C₂₀), suchas 5'- acetyl and 2',3',5'-triacetyl prodrugs (e.g. compounds 42a, 52b,42 and 52a) and pharmaceutical salts such as thiolate, citrate andacetate salts.

The pharmaceutical compositions can be administered topically, orally,or parentally and may take the form of tablets, lozenges, granules,capsules, pills, ampoules or suppositories. They may also take the formof ointments, gels, pastes, creams, sprays, lotions, suspensions,solutions and emulsions of the active ingredient in aqueous ornonaqueous diluents, syrups, granulates or powders. In addition to acompound of the present invention, the pharmaceutical compositions canalso contain other pharmaceutically active compounds or a plurality ofcompounds of the invention.

More particularly, a compound of the formula of the present inventionalso referred to herein as the active ingredient, may be administeredfor therapy by any suitable route including oral, rectal, nasal, topical(including transdermal, buccal and sublingual), vaginal, parental(including subcutaneous, intramuscular, intravenous and intradermal) andpulmonary. It will also be appreciated that the preferred route willvary with the condition and age of the recipient and the nature of theinfection.

In general, a suitable dose for each of the above-named viralinfections, e.g., CMV, EBV, and HHV6 infections, is in the range ofabout 0.1 to about 250 mg per kilogram body weight of the recipient perday, preferably in the range of about 1 to about 100 mg per kilogrambody weight per day and most preferably in the range of about 5 to about20 mg per kilogram body weight per day. Unless otherwise indicated, allweights of active ingredient are calculated as the parent compound ofthe formula of the present invention for salts or esters thereof, theweights would be increased proportionately. The desired dose ispreferably presented as two, three, four, five, six or more sub-dosesadministered at appropriate intervals throughout the day. Thesesub-doses may be administered in unit dosage forms, for example,containing about 10 to about 1000 mg, preferably about 20 to about 500mg, and most preferably about 100 to about 400 mg of active ingredientper unit dosage form. It will be appreciated that appropriate dosages ofthe compounds and compositions of the invention may depend on the typeand severity of the vital infection and can vary from patient topatient. Determining the optimal dosage will generally involve thebalancing of the level of therapeutic benefit against any risk ordeleterious side effects of the antiviral treatments of the presentinvention.

Ideally, the active ingredient should be administered to achieve peakplasma concentrations of the active compound of from about 0.25 to about100 μM, preferably about 0.5 to about 70 μM, most preferably about 1 toabout 50 μM. This may be achieved, for example, by the intravenousinjection of about 0.1 to about 5% solution of the active ingredient,optionally in saline, or orally administered, for example, as a tablet,capsule or syrup containing about 0.1 to about 250 mg per kilogram ofthe active ingredient. Desirable blood levels may be maintained by acontinuous infusion to provide about 0.01 to about 5.0 mg/kg/hour or byintermittent infusions containing about 0.4 to about 15 mg per kilogramof the active ingredient.

While it is possible for the active ingredient to be administered alone,it is preferable to present it as a pharmaceutical formulationcomprising at least one active ingredient, as defined above, togetherwith one or more pharmaceutically acceptable carriers therefor andoptionally other therapeutic agents. Each carrier must be "acceptable"in the sense of being compatible with the other ingredients of theformulation and not injurious to the patient.

Formulations include those suitable for oral, rectal, nasal, topical(including transdermal, buccal and sublingual), vaginal, parenteral(including subcutaneous, intramuscular, intravenous and intradermal) andpulmonary administration. The formulations may conveniently be presentedin unit dosage form and may be prepared by any methods well known in theart of pharmacy. Such methods include the step of bringing intoassociation the active ingredient with the carrier which constitutes oneor more accessory ingredients. In general, the formulations are preparedby uniformly and intimately bringing into association the activeingredient with liquid carriers or finely divided solid carders or both,and then if necessary shaping the product.

Formulations of the present invention suitable for oral administrationmay be presented as discrete units such as capsules, cachets or tablets,each containing a predetermined amount of the active ingredient; as apowder or granules; as a solution or suspension in an aqueous ornon-aqueous liquid; or as an oil-in-water liquid emulsion or awater-in-oil liquid emulsion. The active ingredient may also bepresented a bolus, electuary or paste.

A tablet may be made by compression or moulding, optionally with one ormore accessory ingredients. Compressed tablets may be prepared bycompressing in a suitable machine the active ingredient in afree-flowing form such as a powder or granules, optionally mixed with abinder (e.g., povidone, gelatin, hydroxypropylemthyl cellulose),lubricant, inert diluent, preservative, disintegrant (e.g., sodiumstarch glycollate, cross-linked povidone, cross-linked stadiumcarboxmethyl cellulose) surface-active or dispersing agent. Mouldedtablets may be made by moulding in a suitable machine a mixture of thepowdered compound moistened with an inert liquid diluent. The tabletsmay optionally be coated or scored and may be formulated so as toprovide slow or controlled release of the active ingredient thereinusing, for example, hydroxypropylmethyl cellulose in varying proportionsto provide the desired release profile. Tablets may optionally beprovided with an enteric coating, to provide release in parts of the gutother than the stomach. Formulations suitable for topical administrationin the mouth include lozenges comprising the active ingredient in aflavored basis, usually sucrose and acacia or tragacanth; pastillescomprising the active ingredient in an inert basis such as gelatin andglycerin, or sucrose and acacia; and mouthwashes comprising the activeingredient in a suitable liquid carrier.

Pharmaceutical compositions for topical administration according to thepresent invention may be formulated as an ointment, cream, suspension,lotion, powder, solution, past, gel, spray, aerosol or oil.Alternatively, a formulation may comprise a patch or a dressing such asa bandage or adhesive plaster impregnated with active ingredients andoptionally one or more excipients or diluents.

For infections of the eye or other external tissues, e.g., mouth andskin, the formulations are preferably applied as a topical ointment orcream containing the active ingredient in an amount of, for example,about 0.075 to about 20% w/w, preferably about 0.2 to about 25% w/w andmost preferably about 0.5 to about 10% w/w. When formulated in anointment, the active ingredient may be employed with either a paraffinicor a water-miscible ointment base. Alternatively, the active ingredientsmay be formulated in a cream with an oil-in-water cream base.

If desired, the aqueous phase of the cream base may include, forexample, at least about 30% w/w of a polyhydric alcohol, i.e., analcohol having two or more hydroxyl groups such as propylene glycol,butane-1,3-diol, mannitol, sorbitol, glycerol and polyethylene glycoland mixtures thereof. The topical formulations may desirably include acompound which enhances absorption or penetration of the activeingredient through the skin or other affected areas. Examples of suchdermal penetration enhancers include dimethylsulphoxide and relatedanalogues.

The oily phase of the emulsions of this invention may be constitutedfrom known ingredients in an known manner. While this phase may comprisemerely an emulsifier (otherwise known as an emulgent), it desirablycomprises a mixture of at lease one emulsifier with a fat or an oil orwith both a fat and an oil. Preferably, a hydrophilic emulsifier isincluded together with a lipophilic emulsifier which acts as astabilizer. R is also preferred to include both an oil and a fat.Together, the emulsifier(s) with or without stabilizer(s) make up theso-called emulsifying wax, and the wax together with the oil and/or fatmake up the so-called emulsifying ointment base which forms the oilydispersed phase of the cream formulations.

Emulgents and emulsion stabilizers suitable for use in the formulationof the present invention include Tween 60, Span 80, cetostearyl alcohol,myristyl alcohol, glyceryl monostearate and sodium lauryl sulphate.

The choice of suitable oils or fats for the formulation is based onachieving the desired cosmetic properties, since the solubility of theactive compound in most oils likely to be used in pharmaceuticalemulsion formulations is very low. Thus the cream should preferably be anon-greasy, non-staining and washable product with suitable consistencyto avoid leakage from tubes or other containers. Straight or branchedchain, mono- or dibasic alkyl esters such as di-isoadipate, isocatylstearate, propylene glycol diester of coconut fatty acids, isopropylmyristate, decyl oleate, isopropyl palmitate, butyl stearate,2-ethylhexyl palmitate or a blend of branched chain esters known asCrodamol CAP may be used, the last three being preferred esters. Thesemay be used alone or in combination depending on the propertiesrequired. Alternatively, high melting point lipids such as white softparaffin and/or liquid paraffin or other mineral oils can be used.

Formulations suitable for topical administration to the eye also includeeye drops wherein the active ingredient is dissolved or suspended in asuitable carrier, especially an aqueous solvent for the activeingredient. The active ingredient is preferably present in suchformulation in a concentration of about 0.5 to about 20%, advantageouslyabout 0.5 to about 10% particularly about 1.5% w/w.

Formulations for rectal administration may be presented as a suppositorywith a suitable base comprising, for example, cocoa butter or asalicylate.

Formulations suitable for vaginal administration may be presented aspessaries, tampons, creams, gels, pastes, foams or spray formulationscontaining in addition to the active ingredient, such carriers as areknown in the art to be appropriate.

Formulations suitable for nasal administration, wherein the carrier is asolid, include a coarse powder having a particle size, for example, inthe range of about 20 to about 500 microns which is administered in themanner in which snuff is taken, i.e., by rapid inhalation through thenasal passage from a container of the powder held close up to the nose.Suitable formulations wherein the carrier is a liquid foradministration, for example, a nasal spray or a nasal drops, includeaqueous or oily solutions of the active ingredient.

Formulations suitable for parenteral administration include aqueous andnon-aqueous isotonic sterile injection solutions which may containanti-oxidants, buffers, bacteriostats and solutes which render theformulation isotonic with the blood of the intended recipient; andaqueous and non-aqueous sterile suspensions which may include suspendingagents and thickening agents, and liposomes or other micorparticulatesystems which are designed to target the compound to blood components orone or more organs. The formulations may be presented in unit-dose ormulti-dose sealed containers, for example, ampoules and vials, and maybe stored in a freeze-dried (lyophilized) condition requiring only theaddition of the sterile liquid carrier, for example water forinjections, immediately prior to use. Extemporaneous injection solutionsand suspensions may be prepared from sterile powders, granules andtablets of the kind previously described.

Preferred unit dosage formulations are those containing a daily dose orunit, daily subdose, as herein above-recited, or an appropriate fractionthereof, of an active ingredient.

It should be understood that in addition to the ingredients particularlymentioned above, the formulations of this invention my include otheragents conventional in the art having regard to the type of formulationin question, for example, those suitable of oral administration myinclude such further agents as sweeteners, thickeners and flavoringagents.

Compounds of the formula of the present invention may also be presentedfor the use in the form of veterinary formulations, which may beprepared, for example, by methods that are conventional in the art.

C. Methods of Synthesis

The present invention also comprises methods of synthesis ofpolysubstituted benzimidazoles of the invention. The present inventionprovides a process for the preparation of polysubstituted benzimidazolesof the following formula: ##STR3## wherein R₁ is H, R₂ is Cl, R₃ is Cl,R₄ is H, R₅ is Cl and R₆ is β-D-ribofuranosyl (denoted compound 45 inthe text);

R₁ is H, R₂ is Cl, R₃ is Cl, R₄ is H, R₅ is Br and R₆ isβ-D-ribofuranosyl (denoted compound 52 in the text);

R₁ is H, R₂ is NO₂, R₃ is NO₂, R₄ is H, R₅ is Cl and R₆ isβ-D-ribofuranosyl (denoted compound 61 in the text);

R₁ is Cl, R₂ is H, R₃ is Cl, R₄ is H, R₅ is Cl and R₆ isβ-D-ribofuranosyl (denoted compound 81 in the text);

R₁ is H, R₂ is Cl, R₃ is Cl, R₄ is H, R₅ is I and R₆ isβ-D-ribofuranosyl (denoted compound 83a in the text);

R₁ is Br, R₂ is Br, R₃ is H, R₄ is H, R₅ is Cl and R₆ isβ-D-ribofuranosyl (denoted compound 85 in the text);

R₁ is H, R₂ is Br, R₃ is Cl, R₄ is H, R₅ is Cl and R₆ isβ-D-ribofuranosyl (denoted compound 95 in the text);

R₁ is H, R₂ is Cl, R₃ is Br, R₄ is H, R₅ is Cl and R₆ isβ-D-ribofuranosyl (denoted compound 99 in the text);

R₁ is H, R₂ is I, R₃ is I, R₄ is H, R₅ is Cl and R₆ is β-D-ribofuranosyl(denoted compound 107 in the text);

R₁ is H, R₂ is Cl, R₃ is Cl, R₄ is H, R₅ is Cl and R₆ is2'-deoxy-β-D-erythro-pentofuranosyl (denoted compound 111 in the text);

R₁ is H, R₂ is Cl, R₃ is Cl, R₄ is H, R₅ is Br and R₆ is2'-deoxy-β-D-erythro-pentofuranosyl (denoted compound 112 in the text);

R₁ is H, R₂ is Cl, R₃ is Cl, R₄ is H, R₅ is Br and R₆ is H (denotedcompound 7 in the text);

R₁ is H, R₂ is Cl, R₃ is F, R₄ is H, R₅ is Cl and R₆ is H (denotedcompound 12c in the text);

R₁ is Cl, R₂ is Cl, R₃ is Cl, R₄ is H, R₅ is Cl and R₆ is H (denotedcompound 13 in the text);

R₁ is H, R₂ is NO₂, R₃ is H, R₄ is H, R₅ is Cl and R₆ is H (denotedcompound 19 in the text);

R₁ is H, R₂ is I, R₃ is NO₂, R₄ is H, R₅ is Cl and R₆ is H (denotedcompound 26 in the text);

R₁ is Cl, R₂ is H, R₃ is Cl, R₄ is H, R₅ is Cl and R₆ is H (denotedcompound 32 in the text);

R₁ is H, R₂ is I, R₃ is I, R₄ is H, R₅ is Cl and R₆ is H (denotedcompound 41 in the text);

R₁ is Cl, R₂ is H, R₃ is CF₃, R₄ is H, R₅ is Cl and R₆ is H (denotedcompound 41c in the text);

R₁ is H, R₂ is Cl, R₃ is Cl, R₄ is H, R₅ is NH₂ and R₆ isβ-D-ribofuranosyl (denoted compound 44 in the text);

R₁ is H, R₂ is Cl, R₃ is Cl, R₄ is H, R₅ is SCH₂ C₆ H₅ and R₆ isβ-D-ribofuranosyl (denoted compound 54 in the text);

R₁ is H, R₂ is Br, R₃ is Br, R₄ is H, R₅ is Cl and R₆ isβ-D-ribofuranosyl (denoted compound 57 in the text);

R₁ is H, R₂ is F, R₃ is F, R₄ is H, R₅ is Cl and R₆ is β-D-ribofuranosyl(denoted compound 65 in the text);

R₁ is H, R₂ is Cl, R₃ is F, R₄ is H, R₅ is Cl and R₆ isβ-D-ribofuranosyl (denoted compound 65a in the text);

R₁ is H, R₂ is H, R₃ is Cl, R₄ is H, R₅ is Cl and R₆ isβ-D-ribofuranosyl (denoted compound 67 in the text);

R₁ is Cl, R₂ is H, R₃ is Cl, R₄ is H, R₅ is CF₃ and R₆ isβ-D-ribofuranosyl (denoted compound 81b in the text);

R₁ is Cl, R₂ is H, R₃ is CF₃, R₄ is H, R₅ is Cl and R₆ isβ-D-ribofuranosyl (denoted compound 81c in the text);

R₁ is H, R₂ is Br, R₃ is H, R₄ is H, R₅ is Cl and R₆ isβ-D-ribofuranosyl (denoted compound 87 in the text);

R₁ is H, R₂ is H, R₃ is Br, R₄ is H, R₅ is Cl and R₆ isβ-D-ribofuranosyl (denoted compound 90 in the text);

R₁ is Cl, R₂ is Cl, R₃ is Cl, R₄ is H, R₅ is Cl and R₆ isβ-D-ribofuranosyl (denoted compound 92 in the text);

R₁ is Br, R₂ is Br, R₃ is Br, R₄ is H, R₅ is Cl and R₆ isβ-D-ribofuranosyl (denoted compound 103 in the text);

R₁ is H, R₂ is Cl, R₃ is Cl, R₄ is H, R₅ is NH₂ and R₆ is2'-deoxy-β-D-erythro-pentofuranosyl (denoted compound 113 in the text);

R₁ is H, R₂ is Cl, R₃ is Cl, R₄ is H, R₅ is Cl and R₆ isβ-D-arabinofuranosyl (denoted compound 134 in the text);

R₁ is Cl, R₂ is Cl, R₃ is Cl, R₄ is Cl, R₅ is Cl and R₆ is(1,3-dihydroxy-2-propoxy)methyl (denoted compound 155 in the text);

R₁ is Cl, R₂ is Cl, R₃ is Cl, R₄ is Cl, R₅ is NH₂ and R₆ is(1,3-dihydroxy-2-propoxy)methyl (denoted compound 156 in the text);

R₁ is Cl, R₂ is Cl, R₃ is Cl, R₄ is Cl, R₅ is Cl and R₆ is2-hydroxyethoxymethyl (denoted compound 166 in the text);

R₁ is Cl, R₂ is Cl, R₃ is Cl, R₄ is Cl, R₅ is OCH₃ and R₆ is2-hydroxyethoxymethyl (denoted compound 166a in the text);

R₁ is Cl, R₂ is Cl, R₃ is Cl, R₄ is Cl, R₅ is NH₂ and R₆ is2-hydroxyethoxymethyl (denoted compound 167 in the text);

R₁ is H, R₂ is Cl, R₃ is Cl, R₄ is H, R₅ is Cl and R₆ is5-O-acetyl-β-D-ribofuranosyl (denoted compound 42a in the text);

R₁ is H, R₂ is Cl, R₃ is Cl, R₄ is H, R₅ is Br and R₆ is5-O-acetyl-β-D-ribofuranosyl (denoted compound 52b in the text);

R₁ is H, R₂ is Cl, R₃ is Cl, R₄ is H, R₅ is Cl and R₆ is2,3,5-tri-O-acetyl-β-D-ribofuranosyl (denoted compound 42 in the text);

R₁ is H, R₂ is Cl, R₃ is Cl, R₄ is H, R₅ is Br and R₆ is2,3,5-tri-O-acetyl-β-D-ribofuranosyl (denoted compound 52a in the text);

which comprises reacting a compound of the following formula: ##STR4##wherein R₁, R₂, R₃, R₄ and R₅ are defined as above, with a protectedcompound of formula R₆ ^(a) X, shown below, in which R₆ ^(a) representsa protected carbohydrate or carbohydrate-like moiety and X represents aleaving group, and subsequently removing the protecting group from themoiety to form the desired compound.

The compound of formula R₆ ^(a) X is represented by the formula:##STR5## wherein X represents a leaving group such as a halo (e.g.chloro) or C₁₋₄ alkanoyloxy (e.g. acetoxy) group and R represents ahydroxy protecting group, for example, an acyl group such as C₁₋₄alkanoyl (e.g. acetyl or aroyl, e.g. benzoyl, e.g. p-nitrobenzoyl andthe like.

The reaction of the compound of formula II with the compound of formulaR₆ ^(a) X may be effected in the presence of BSA, TMSOTf, BSTFA and thelike, advantageously in a solvent such as acetonitrile, dichloroethaneand the like.

The removal of the protecting groups in the above process according tothe invention may be effected, for example, by treatment with methanolicammonia, boron trihalide, sodium carbonate, potassium cyanide and thelike.

Additionally, compounds of formula II may be reacted with compounds ofthe formula:

    R'.sup.˜ X

wherein R'=aralkyl

to prepare a compound of general formula I, wherein R₆ is benzyl ratherthan a carbohydrate or carbohydrate-like moiety, and wherein, e.g. R₁ isH, R₂, R₃ is Cl, R₄ is H and R₅ is NH₂ or Cl.

The compounds of the present invention, can be synthesized in accordancewith the representative procedures described vide infra. In general, theappropriate benzimidazole was prepared and then condensed with theappropriate precursor for the ultimate R₆ group as represented by theschemes and substituent charts of FIGS. 1 and 2. This furnishedpolysubstituted benzimidazoles which had potential antiviral activity,and were also amenable toward subsequent chemical transformations toafford additional compounds. The solvents, reagents and reactionconditions for the preparation of some representative startingmaterials, intermediate and target compounds are presented in detailbelow. The compound numbering in this section and specification refersto the reaction scheme and chart numbers of the compounds.

SPEClFIC EXAMPLES 2,5,6-Trichlorobenzimidazole(5)

Method A(5)

A solution of 5,6-dichlorobenzimidazole-2-one (5 g, 2.5 mmole) in 75 mlof POCl₃ was heated at reflux for 5 hr. HCl gas was passed into themixture for the last 1/2 hr. Excess POCl₃ was removed in vacuo and theresidue was decomposed with HCl (150 ml). The brown solid was removed byfiltration and washed with H₂ O (100 ml). The filtrate was made slowlybasic with NH₄ OH. After cooling, the precipitate was filtered, theprecipitate was dissolved in MeOH and an insoluble material was removedby filtration. The filtrate was evaporated under diminished pressure todryness. The residue was applied to a column of silica gel (Kiesel Gel70-230 mesh) and eluted with CH₂ Cl₂. The fractions containing theproduct (5) were combined, and evaporated to dryness. Yield 1.36 g(25.0%).

Method B(5)

A CuCl₂ saturated aqueous solution (15 ml) was diluted to 25 ml withwater. Sodium nitrite (1.035 g, 5 mmole) was dissolved in 5 ml of waterand slowly added to the CuCl₂ solution. After two minutes,2-amino-5,6-dichlorobenzimidazole(4) (0.935 g, 5 mmole) was slowly addedin small portions. The mixture was stirred at room temperature for 1 hr.Excess CuCl₂ solution was added and the mixture was heated on a steambath for 1 hr. The aqueous solution was then extracted with ethylacetate (3×50 ml) and the organic layer was washed with brine, driedwith MgSO₄, concentrated, and separated on a silica column using 2%MeOH/CHCl₃ to afford 545 mg (49.5%) of 2,5,6-trichlorobenzimidazole(5).¹ H NMR, TLC, and MS analysis were identical to the same compoundobtained by Method A.

Method C(5)

5,6-Dichlorobenzimidazole-2-thione (3) was prepared according to themethod of Van Allen and Deacon, as described in Van Allen, J. A. et al.,Org. Syn. IV:569. Carbon disulfide (84.5 mL 107.0 g, 1.4M) was added toa solution of KOH (92.4 g, of 85% purity, 78.5 g, 1.4M) in EtOH (1.5 L)and the resulting yellow solution was stirred without heating until awhite precipitate was obtained. To obtain the product in good yield,potassium ethyl xanthate must be prepared in situ.4,5-Dichloro-1,2-phenylenediamine (201.8 g, 1.14M) dissolved in absoluteEtOH (3 L) was then added and the reaction mixture was heated underreflux for 24 hr. After cooling to room temperature the solvent wasremoved (caution--stench) under reduced pressure and the residuedissolved in H₂ O (3.0 L). The solution was adjusted to pH 2 with AcOHto precipitate the product. This solid was collected by filtration,washed with H₂ O, and the solid was dried under reduced pressure at 60°C. for 48 hr to yield 236.4 g (94.6%) of5,6-dichlorobenzimidazole-2-thione (3), MP: 328°-330° C. Alsocommercially available from Maybridge Chemical Company, Ltd., lit. MPgiven as >320° C. TLC: R_(f) =0.62 (CHCl₃ -MeOH, 10:1; SiO₂).

To a five liter three neck roundbottom flask fitted with an overheadstirrer and a reflux condenser was added H₂ O (1500 mL) and sodiumpercarbonate commercially available from Fluka Corporation. (209.3 g,1.2M, 90% pure). For a leading reference see Ando, T. et al., Chem.Letters 665-666 (1986). After a solution was obtained,5,6-dichlorobenzimidazole-2-thione (3) (131.5 g, 0.6M) was added inseveral portions. (Caution: exothermic with foaming). After stirringovernight, without heating, an additional 21 g of sodium percarbonatewas added and the mixture was heated under reflux until a clear solutionwas obtained. The hot solution was treated with charcoal, filteredthrough Celite, then cooled to 25° C. in an ice bath. With cooling,concentrated HCl was added very carefully until pH 1 was obtained. Theproduct precipitated as a white crystalline solid. After storing at 5°C. for 24 hr, the precipitate was collected by filtration, washed withcold H₂ O (500 mL) then dried at 80° C. under reduced pressure for 20 hrto yield 135.0 g (84%) of 5,6-dichlorobenzimidazole-2-sulfonic acid (9).MP: 353°-354° C. (eff.). TLC: R_(f) =0.27 (CHCl₃ -MeOH, 4:1, SiO₂).

A modified procedure of Balli and Kersting procedure described in Balli,H. et al., Justis Liebigs Am. Chem, 1:647 (1961), was followed. A oneliter three neck roundbottom flask was fitted with an over head stirrerand a reflux condenser connected to a gas scrubber. To the flask wasadded in order, phosphorus oxychloride (80 mL), phosphorus pentachloride(104.0 g, 0.5M) and 5,6-dichlorobenzimidazole-2-sulfonic acid (9) (66.8g, 0.25M). The resulting mixture was carefully heated until anexothermic reaction occurred. The heat source was removed. When thereaction had subsided, the mixture was carefully heated to reflux. Afterall gas evolution had ceased (5 hr) the mixture was protected frommoisture and allowed to cool to 25° C. The resulting thick slurry wascarefully added to cold H₂ O (2.5 L) containing some ice. Ice was addedas needed. The aqueous mixture was allowed to stand for 18 hr at 5° C.while the product precipitated. The pH of the mixture was adjusted to pH8 with conc. NH₄ OH then acidified to pH 3 with glacial AcOH. The solidwas collected by filtration, washed with H₂ O, then dried on the filter.The material was dissolved in THF, treated with charcoal, then filteredthrough Celite. The solvent was removed under reduced pressure. Thesolid residue was dried at 60° C. under reduced pressure for 18 hr toyield 39.9 g (72%) of 2,5,6-trichlorobenzimidazole (5). MP: 212°-214° C.(eff. then solidifies). See Kawashima, E. et al., Nucleic AcidChemistry: Improved and New Synthetic Procedures, Methods andTechniques, eds. Townsend, L. B. et al., Wiley Interscience New York,N.Y., Part IV p. 96 (1991), TLC: R_(f) =0.89 (EtOAc; SiO₂).Recrystallization from benzene-hexane raises the melting point to223°-224° C.

2-Amino-5,6-dichlorobenzimidazole (4)

A modified procedure of the procedure described by Leonard, N. J., etal., J. Am. Chem. Soc. 69:2459 (1947), was followed. Cyanogen bromide(136.6 g, 1.3M, 260 mL of a 5M solution in CH₃ CN from) was added to asolution of MeOH (250 mL) in H₂ O (1500 mL). 4,5-Dichloro-1,2-phenylenediamine (available commercially from AldrichChemical Company) (222.4 g, 1.26M) was then added in five portions, asthe initial reaction is exothermic. The reaction mixture was stirredwithout heating for 80 hr. then treated with activated charcoal (5 g).The 80 hr time period was for convenience alone, since the reaction isprobably complete within 24 hr. After stirring for two hrs, the reactionmixture was filtered through Celite. The filter cake was washed withMeOH (250 mL) and H₂ O (750 mL) and the filter cake was discarded. Thefiltrate was diluted with H₂ O (1.5 L), adjusted to pH 10 with conc. NH₄OH (1750 mL) and then allowed to stand overnight at 5° C. Theprecipitate (yellow leaflets) was collected by filtration, washed withH₂ O and then dried under reduced pressure at 50° C. for 60 hr. Crudeyield: 265 g. This product was purified by recrystallization from CH₃ CNto yield: 205.6 g (80.7%) of 4. MP: 264°-266° C. TLC: R_(f) =0.22 (CHCl₃-MeOH), 10:1; SiO₂). See Homer, J. K. et al., J. Med. Chem. 11:946-949(1968), lit. MP: 260°-262° C. As an alternative, the crude material isdissolved in absolute ethanol, treated with activated charcoal, and thesolution diluted with an equal volume of H₂ O and stored at 5° C. for18-24 hr.

2-Bromo-5,6-dichlorobenzimidazole (7)

Method A(7)

2-Amino-5,6-dichlorobenzimidazole (3 g, 16 mmole) was suspended in 150ml of water and brought into solution with 2 ml of HBr. Sodium nitrite(3.3 g, 55 mmole) was then added and the mixture was stirred at roomtemperature for 1 hr. Excess CuBr₂ was then added and the mixture washeated on a steam bath for 1 hr. The aqueous solution was extracted withethyl acetate (3×100 ml), dried with MgSO₄, concentrated, andcrystallized from ethyl ether to give 1.13 g (26%) of2-bromo-5,6-dichlorobenzimidazole (7). ¹ H NMR (DMSO-d₆) δ 7.81 ppm (s,2H), 13.62 (s, 1H). GC/MS: m/e 266, 185, 158, 150, 133, 124, 107, 97,88, 73, 62, 52, 37.

Method B(7)

This preparation was based on work reported by Doyle et al., J. Org.Chem. 42:2426 (1977). A two liter three neck roundbottom flask wasfitted with an overhead stirrer and a reflux condenser with an attachedbubbler. Acetone (225 mL), that had been filtered through anhydrousmagnesium sulfate, and isopentyl nitrite (25.1 mL, 29.2 g, 0.25M) wereadded and the resulting solution stirred for 5 min. CuBr₂ (23.5 g,0.105M) was then added and stirring was continued for an additional 75min to give a yellow-black solution. Powdered2-amino-5,6-dichlorobenzimidazole (3) (20.2 g, 0.1M) was added in oneportion and the mixture stirred without heating. Gas evolution wasalmost immediate and very vigorous; the reaction was exothermic to thepoint that a gentle reflux was maintained. During this time, an orangesolid began to separate. After 25 min, the rate of reflux began todecrease, and after 35 min was very slow. The mixture was then heatedunder reflux until gas evolution had almost ceased. The mixture wascooled for 20 min in an ice bath then treated with 48% aqueous HBr (100mL) giving a black suspension. After stirring for 20 min, H₂ O was addedand an orange precipitate was obtained. Stirring was continued for 30min and then the precipitate was removed by filtration. (Caution--stronglachrymator, probably bromoacetone, present). The solid was suspended inH₂ O and the mixture adjusted to pH 7 with saturated NaHCO₃. The solidwas collected by filtration washed with H₂ O then dried at 50° C. underreduced pressure for 18 hr to yield 20.9 g (79%) of crude (7). TLC:R_(f) =0.0, 0.36 (5,6-dichlorobenzimidazol-2-one), 0.80 (7)(EtOAc-hexanes, 5:1; SiO₂). TLC: R_(f) =EtOAc-hexanes, 5:1; SiO₂). NMR:(DMSO-d₆) d 7.804, s, 2H, H4, H7. UV (EtOH): pH7 224, 253, 260, 292,299; pH1 211, 251, 289, 298; pH11 221, 294, 301.

2,5,6-Trichloro-1-(2,3,5-tri-O-acetyl-β-D-ribofuranosyl)benzimidazole(42)

To a suspension of (5) (62.013 g, 280 mmol) in 1.4 L of dry MeCN underargon, was added 70 mL (280 mmol) of BSA. The reaction mixture wasstirred at room temperature for 15 min to give a clear solution. To thissolution were added 1,2,3,5-Tetra-O-acetyl-β-D-ribofuranose (89.096 g,280 mmol) and then 60 mL (310 mmol) of TMSOTf was added dropwise over 20min. After the addition had been completed, stirring was continued atroom temperature for an additional 30 min. The reaction mixture wasdiluted with 4 L of EtOAc. The EtOAc solution was washed with 1:1saturated aqueous NaHCO₃ /saturated aquenous NaCl (3×2 L), dried (withNa₂ SO₄, 100 g), decolorized (activated charcoal, 8 g) and filteredthrough Celite. The filtrate was evaporated and the residue wasrecrystallized from MeOH to give 116.88 g (3 crops, 87%) of (42) aswhite needles. The recrystallization was accomplished by heating thecrude product in MeOH at reflux on a steam bath for a few minutes andthen decanting the supernatant while hot into a clean beaker. Immediatecrystallization resulted. A second portion of fresh MeOH was added tothe remaining solid and the whole process was repeated until all of thesolid was dissolved. MP: 145°-146° C. This product was pure by ¹ H NMR.

2,5,6-Trichloro-1-(β-D-ribofuranosyl)benzimidazole (45)

Method A(45)

2,5,6-Trichlorobenzimidazole (700 mg, 0.0032 moles) was dissolved inacetonitrile and BSTFA (1 ml, 0.0038 moles) was added. The mixture washeated at 75° C. for 20 minutes. TMSTf (1 ml, 0.0051 moles) and1-O-acetyl-2,3,5-tri-O-benzoyl-β-D-ribofuranose (1.9 g, 0.0038 moles)were added while heating was continued for 45 min. The acetonitrile wasremoved under reduced pressure and the protected nucleoside wasseparated on a silica column, eluting with chloroform. The benzoylprotecting groups were removed by overnight treatment at roomtemperature with methanolic ammonia. The nucleoside was separated on acolumn using 50% EtOAc/hexane and then 10% MeOH/CHCl₃. The isolatedcompound was recrystallized from methanol. Yield: (74%); MP: 185°-186°C.; TLC (10% MeOH/CHCl₃): R_(f) =0.20; ¹ H NMR (DMSO-d₆) δ 3.68 ppm (q,2H), 4.00 (d, 1H), 4.12 (t, 1H), 4.40 (q, 1H), 5.28 (d, 1H), 5.41 (t, 1H), 5.49 (d, 1H), 5.57 (d, 1H), 7.96 (s, 1H), 8.55 (s, 1H); ¹³ C NMR(DMSO-d₆, Broad band decoupling): d 61.08 ppm, 69.80, 71.70, 86.47,89.16, 114.93, 120.04, 125.77, 125.97, 132.30, 140.96, 142.16; MS (FastAtom Bombardment): m/e (M⁺ 6) 359, (M⁺ 4) 357, (M⁺ 2) 355, (M⁺) 353,319, 285, 263, 221, 187, 177, 133, 115, 103, 97, 85.

Method B(45)

To a solution of Na₂ CO₃ (23.32 g, 220 mmol) in 440 mL of H₂ O wereadded successively 2.0 L of EtOH, 2.0 L of MeOH, and 105.5 g (220 mmol)of 42. The reaction mixture was stirred at room temperature for 2 hr.AcOH (26.44 mL 462 mmol) was added and stirring was continued at roomtemperature for 20 min. The reaction mixture was filtered. The solidproduct was triturated with H₂ O (800 mL 20 min), MeOH (500 mL 30 min),and then recrystallized from EtOH/MeOH (1 L/1 L) to give 52.4 g of 45 aswhite crystals {2 crops, the 2nd crop (2.8 g) was obtained byevaporation of the mother liquid and recrystallization of the residuetwo more times from EtOH/MeOH}. Recrystallization was accomplished bydissolving the crude product in a minimum amount of boiling EtOH,diluting it immediately with an equal amount of hot MeOH and allowing itto slowly cool. The filtrate was evaporated and the residue wastriturated with H₂ O (500 mL, 20 min), MeOH (500 mL, 30 min), and thenrecrystallized twice from EtOH/MeOH to give an additional 15.34 g of 45{2 crops, the 2nd crop (1.95 g) was obtained by evaporation of themother liquid and recrystallization of the residue two more times fromEtOH/MeOH}. The total of 45 was 67.74 g (87%). MP: 166°-168° C.

2-Bromo-5,6-dichloro-1-(2,3,5-tri-O-acetyl-β-D-ribofuranosyl)benzimidazole(52a)

To a suspension of (7) (47.867 g, 180 mmol) in 540 mL of dry. MeCN, wasadded 45 mL (180 mmol) of BSA. The reaction mixture was stirred at roomtemperature for 15 min to give a clear solution. A small amount of solidimpurity remained unsilylated, believed to be5,6-dichlorobenzimidazol-2-one from the starting material(7).1,2,3,5-Tetra-O-acetyl-β-D-ribofuranose (60.14 g, 189 mmole) was addedto this solution and then TMSOTf (38.267 mL 198 mmol) was added dropwiseover 20 min. After the addition had been completed, stirring wascontinued at room temperature for 30 min. The reaction mixture wasdiluted with 2 L of EtOAc. The EtOAc solution was washed with 1:1saturated aquenous NaHCO₃ /saturated aquenous NaCl (1.5 L×2), dried (Na₂SO₄, 100 g), decolorized (activated charcoal, 6 g), and filtered throughCelite. The filtrate was evaporated and the residue was recrystallizedfrom MeOH to give 82.21 g (4 crops, 87%) of 52a as white needles. MP:140°-144° C. This product was pure by ¹ H NMR.

2-Bromo-5,6-dichloro-1-(β-D-ribofuranosyl)benzimidazole (52)

Method A(52)

2-Bromo-5,6-dichlorobenzimidazole (7) (1 g, 3.8 mmole) was dissolved indry acetonitrile (150 ml) and stirred in an inert atmosphere at 60° C.BSA (1.03 ml, 4.2 mmole) was added and the mixture was stirred for 10minutes. 1,2,3,5-Tetra-O-acetyl-β-D-ribofuranose (1.21 g, 3.8 mmole) andTMSTf (0.81 ml, 4.2 mmole) were added to the clear solution and themixture was stirred for 10 min. An additional quantity of1,2,3,5-tetra-O-acetyl-D-ribofuranose (1.21 g, 3.8 mmole) and TMSTf(0.81 ml, 4.2 mmole) were added to the clear solution and the mixturewas allowed to stir at 6° C. for 1 hr. The mixture was concentratedunder reduced pressure and separated on a silica column to give theprotected 2-bromo-5,6-dichlorobenzimidazole nucleoside. ¹³ C NMR(CDCl₃): δ 170.26 ppm, 169.50, 168.99, 142.97, 132.32, 130.71, 128.09,128.06, 121.09, 112.98, 88.09, 80.76, 71.01, 69.50, 62.87, 20.97, 20.49,20.13. ¹ H NMR (CDCl₃): δ 2.02 ppm (s, 3H), 2.16 (s, 3H), 2.29 (s, 3H),4.38 (m, 1H), 4.46 (ddd, 2H), 5.43 (dd, 1H), 5.43 (t, 1H), 6.17 (d, 1H),7.78 (s, 1H), 7.81 (s, 1H). The protected nucleoside was stirredovernight at room temperature in a methanolic ammonia solution,concentrated, and suspended in methanol (3×25 ml) to yield2-bromo-5,6-dichloro-1-(β-D-ribofuranosyl)benzimidazole nucleoside in137%. ¹³ C NMR (DMSO-d₆): δ 142.57 ppm, 132.60, 132.57, 125.76, 119.86,114.73, 90.21, 86.35, 71.55, 69.76, 61.05. ¹ H NMR (DMSO-d₆): δ 3.69 ppm(m, 2H), 3.99 (m, 1H), 4.11 (m, 1H), 4.41 (q, 1H), 5.27 (d, 1H), 5.40(t, 1H), 5.45 (d, 1H), 5.87 (d, 1H), 7.95 (s, 1H), 8.56 (s, 1H). MS(FAB): m/e 399, 351, 319, 285, 267, 219, 187, 153, 133, 103, 85.

METHOD B (52)

To a solution of Na₂ CO₃ (2.12 g, 20 min) in 40 mL of H₂ O, were addedsuccessively 180 mL of EtOH, 180 mL of MeOH, and 10.483 g (20 mmol) of51a. The reaction mixture was stirred at room temperature for 1.5 hr.AcOH (2.404 mL, 42 mmol) was added and stirring was continued at roomtemperature for 10 min. The reaction mixture was evaporated and thesolid product was triturated with H₂ O (200 mL, 20 min), MeOH (100 mL,30 min), and then recrystallized from EtOH/MeOH, by dissolving the crudeproduct in a minimum amount of boiling EtOH, diluting it immediatelywith an equal amount of hot MeOH and allowing it to slowly cool, to give6.87 g of 52 as white crystals {2 crops, the 2nd crop (0.609 g) wasobtained by evaporation of the mother liquor and recrystallization ofthe residue two more times from EtOH/MeOH}. MP: 162°--162° C.

2-Chloro-5,6-dinitro-1-(2,3,5-tri-O-benzyl-β-D-ribofuranosyl)benzimidazole(60)

To a mixture of 0.541 g (2.23 mmol) of 2-chloro-5,6-dinitrobenzimidazolein 12 mL of MeCN, was added 0.558 mL (2.23 mmol) of BSA. The reactionmixture was stirred at 75° C. for 15 min to give a clear solution. Thissolution was treated with the above MeCN solution of2,3,5-tri-O-benzyl-D-ribofuranosyl chloride and 0.56 mL (2.90 mmol) ofTMSOTf at 75° C. for 30 min. The reaction mixture was cooled and dilutedwith EtOAc (50 mL). The EtOAc solution was washed with sat. NaHCO₃solution (50 mL×2), sat. NaCl solution (50 mL), dried (Na₂ SO₄), andevaporated. The residue was chromatographed on a silica column (2.2×18cm, eluted with 20% EtOAc/hexane). Evaporation of fractions 10-14 (20 mLper fraction) gave 0.502 g (35%) of 60 as a syrup. MS: (FAB) m/e 645(1%, MH⁺ =645). ¹ H NMR (DMSO-d₆): δ8.50, 8.49 (2×s, 2, 7-H and 4-H),7.34, 6.90 (2×m, 15, 3×Ph), 6.09 (d, 1, 1'-H, J_(1'-2') =8.0 Hz),4.73˜4.27 (m, 9, 2'-H, 3'-H, 4'-H, and 3×PhCH₂), 3.77 (dd, 1, 5'-H,J_(4'-5') =2.0 Hz, J_(5'-5") =11.0 Hz), 3.65 (dd, 1, 5"-H, J_(4'-5")=2.5 Hz).

2-Chloro-5,6-dinitro-1-β-D-ribofuranosylbenzimidazole (61)

To a solution of 0.464 g (0.719 mmol) of 60 in 12 mL of CH₂ Cl₂, wasadded dropwise 8.4 mL of 1M BCl₃ at -78° C. The reaction mixture wasstirred at -78° C. for 2 hr and then at -40° C. for 2 hr. MeOH (5 mL)was added and stirring was continued at -40° C. for 10 min. The reactionmixture was diluted with EtOAc (75 mL). The EtOAc solution was washedwith cold H₂ O (50 mL), sat. NaHCO₃ solution (50 mL), sat. NaCl solution(50 mL), dried (Na₂ SO₄), and evaporated. The residue was coevaporatedwith MeOH (3×) and then suspended in a small amount of CHCl₃ for a fewhours. The solid product was filtered to give 0.209 g of 61. This samplewas contaminated by a small amount of 2-chloro-5,6-dinitrobenzimidazole.A part of the sample (0.18 g) was purified on a silica column (2.4×10cm, eluted with pure EtOAc). Evaporation of the appropriate fractionsand crystallization by addition of CHCl₃ gave 0.084 g of 61 as a whiteSolid. MP: 132°-135° C. MS: (El) m/e 374.0276 (1%, M⁺ =374.0265). ¹ HNMR (DMSO-d₆): δ 9.18 (s, 1, 7-H), 8.60 (s, 1, 4-H), 6.00 (d, 1, 1'-H,J_(1'-2') =7.5 Hz), 5.59 (d, 1, 2'-OH, J_(2'-2'OH) =6.0 Hz), 5.52 (t, 1,5'-OH, J_(5'-5'OH) =4.5 Hz), 5.38 (d, 1, 3'-OH, J_(3'-3'OH) =4.5 Hz),4.01 (m 1, 2'-H, J_(2'-3') =5.5 Hz), 4.16 (m, 1, 3'-H, J_(3'-4') =2.0Hz), 4.07 (m, 1, 4'-H, J_(4'-5') =J_(4'-5") =2.5 Hz), 3.73 (m, 2, 5'-Hand 5"-H, J_(5'-5") =12.0 Hz). ¹³ C NMR (DMSO-d₆): δ 146.55 (C2), 142.63(C3a), 138.90, 138.60 (C5 and C6), 134.12 (C7a), 116.67 (C4), 111.62(C7), 89.98 (C1'), 86.91 (C4'), 72.86 (C2'). 69.84 (C3'), 60.85 (C5').Anal Calcd. for C₁₂ H₁₁ ClN₄ O₈ : C, 38.47; H, 2.96; N, 14.95. Found: C,38.46; H, 2.98; N, 14.62.

1-(2,3,5-Tri-O-acetyl-β-D-ribofuranosyl)-2,4,6-trichlorobenzimidazole(80)

To a suspension of 1.362 g (6.15 mmol) of 2,4,6-trichlorobenzimidzole in31 mL of MeCN, was added 1.52 mL (6.15 mmol) of BSA. The reactionmixture was stirred at 80° C. for 15 min to give a clear solution. Thissolution was treated with 2.153 g (6.675 mmol) of1,2,3,5-tetra-O-acetyl-β-D-ribofuranose and 1.426 mL (7.380 mmol) ofTMSOTf at 80° C. for 1 hr. The reaction mixture was cooled and dilutedwith EtOAc (150 mL). The EtOAc solution was washed with sat. NaHCO₃solution (150 mL×2), sat. NaCl solution (150 mL), dried (Na₂ SO₄), andevaporated. The residue was coevaporated with MeOH and then suspended in50 mL of hot MeOH. The suspension was cooled, filtered, and the solidproduct was washed with MeOH to give 2.103 g of 80 as white crystals(This product showed one spot on TLC). The mother liquor was evaporatedand the residue was chromatographed on a silica column (2.5×20 cm,eluted with CHCl₃ and 0.5% MeOH/CHCl₃). Evaporation of fractions 20-24(20 mL per fraction) and recrystallization from MeOH gave an additional0.258 g of 80 as white crystals. The total yield of 80 was 2.361 g(80%). MP: 198°-200° C. MS: (El) m/e 480.0069 (4%, for C₁₈ H₁₇ ³⁵ Cl₂ ³⁷ClN₂ O₇ : M⁺ =480.0072). ¹ H NMR (DMSO-d₆): δ 7.89 (d, 1, 7-H, J₇₋₅ =2.0Hz), 7.58 (d, 1, 5-H), 6.26 (d, 1, 1'-H, J_(1'-2') =7.0 Hz), 5.54 (t, 1,2'-H, J_(2'-3') =7.0 Hz), 5.44 (dd, 1, 3'-H, J_(3'-4') =4.5 Hz), 4.48,4.38 (2×m, 3, 4'-H and 5'-H), 2.14, 2.02 (2×s, 9, 3×Ac). ¹³ H NMR(DMSO-d₆): δ 169.84, 169.37, 169.09 (3×COCH₃), 140.91 (C2), 137.39(C3a), 134.14 (C7a), 128.47 (C6), 123.66 (C4), 123.25 (C5), 111.06 (C7),86.91 (C1'), 79.58 (C4'), 70.54 (C2'), 58.58 (C3'), 62.47 (C5'), 20.48,20.19, 19.90 (3×COCH₃). Anal. Calcd. for C₁₈ H₁₇ Cl₃ N₂ O₇ : C, 45.07;H, 3.57; N, 5.34. Found: C, 45.08; H, 3.62; N, 5.87.

2,4,6-Trichloro-1-β-D-ribofuranosylbenzimidazole (81)

A mixture of 0.130 g (0.271 mmol) of 80 in 5 mL of conc. NH₄ OH/dioxane(1:1 by volume) was stirred in a pressure bottle at room temperature for1 day. Volatile materials were removed by evaporation and coevaporationwith MeOH (3×, bath temperature <40° C.). The resulting solid wasabsorbed on silica gel and was chromatographed on a silica column (2×5cm, eluted successively with 1%, 2%; 3% MeOH/CHCl₃). Evaporation offractions 23-39 (5 mL per fraction) gave a white solid. This solid waswashed with H₂ O, dried to give 46 mg (48%) of 80 as a white solid. MP:165°-167° C. ¹ H NMR (DMSO-d₆): δ 8.36 (d, 1, 7-H, J₇₋₅ =2.0 Hz), 7.52(d, 1, 5-H), 5.90 (d, 1, 1'-H, J_(1'-2') =8.0 Hz), 5.49 (d, 1, 2'-OH,J_(2'-2'OH) =6.5 Hz), 5.38 (t, 1, 5'-OH, J_(5'-5'OH) =4.5 Hz), 5.28 (d,1, 3'-OH, J_(3'-3'OH) =4.5 Hz), 4.41 (m, 1, 2'-H, J_(2'-3') =5.5 Hz),4.14 (m, 1, 3'-H, J_(3'-4') =2.0 Hz), 4.02 (m, 1, 4'-H, J_(4'-5')=J_(4'-5") =2.5 Hz), 3.70 (m, 2, 5'-H and 5"-H, J_(5'-5") =12.0 Hz). ¹³H NMR (DMSO-d₆): δ 141.64 (C2), 137.50 (C3a), 134.17 (C7a), 128.02 (C6),123.22 (C4), 122.77 (C5), 112.56 (C7), 89.35 (C1'), 86.52 (C4'), 71.71(C2'), 69.70 (C3'), 61.02 (C5'), Anal. Calcd. for C₁₂ H₁₁ Cl₃ N₂ O₄ : C,40.76; H, 3.14; N, 7.92. Found: C, 40.74; H, 3.37; N, 7.71.

2-Iodo-5,6-dichloro-1-β-D-ribofuranosylbenzimidazole (83a)

2-Amino-5,6-dichloro-1-(2,3,5-tri-O-acetyl-β-D-ribofuranosyl)benzimidazolenucleoside (43) (50 mg, 0.1 mmole) was added to 3 mL of diiodomethaneand amyl nitrite (0.13 ml, 1 mmole) under an inert atmosphere and heatedto 50° C. for 1 hr. The mixture was concentrated under reduced pressureand chromatographed on a silica gel column to give one product which wastreated with methanolic ammonia for 18 hr. The product was isolated,recrystallized and characterized as compound 83a. ¹ H NMR (DMSO-d₆): δ3.71 (m, 2H), 4.07 (m, 1H), 4.12 (m, 1H), 4.40 (q, 1H), 5.23 (m, 1H),5.37 (m, 2H) 5.82 (d, 1H), 7.89 (s, 1H), 8.52 (s, 1H). ¹³ C NMR(DMSO-d₆): δ 145.33 ppm, 132.51, 125.26, 125.18, 119.51, 114.22, 110.32,92.11, 86.22, 71.47, 69.99, 61.11. MP: 180°-182° C. UV λ_(max) nm(ε×10⁴): (pH 7) 230 (.490), 259 (.216), 292 (.277) 302 (.305); (pH 1)230 (.342), 258 (.095), 294 (.301), 303 (.313); (pH 11) 228 (.560), 258(.198), 292 (.249), 302 (.267). Anal. Calcd. for C₁₂ H₁₁ Cl₂ lN₂ O₄1.5CH₃ OH: C, 32.88; H, 3.47; N, 5.68. Found: C, 33.32; H, 3.21; N,5.80.

2-Chloro-4,5-dibromo-1-(2,3,5-tri-O-acetyl-β-D-ribofuranosyl)benzimidazole(84)

To a stirred mixture of 1.228 g (5.498 mmol) of CuBr₂ and 0.654 mL(4.949 mmol) of 90% t-BuONO in 10 mL of CH₃ CN, was added dropwise asolution of 1.170 g (2.748 mmol) of5-amino-2-chloro-1-(2,3,5-tri-O-acetyl-β-D-ribofuranosyl)benzimidazolein 3 mL of CH₃ CN. After the addition, stirring was continued at roomtemperature for 2 hr. The reaction mixture was diluted with 100 mL ofEtOAc. The EtOAc solution was washed with H₂ O (100 mL), sat. NaHCO₃solution (100 mL×2), sat. NaCl solution (100 mL), dried (Na₂ SO₄), andevaporated. The residue was chromatographed on a silica column (4.1×30cm, eluted with CHCl₃). Evaporation of fractions 59-78 (20 mL perfraction) and recrystallization from MeOH gave 0.600 g (38%) of 84 as awhite solid. MP 202°-203° C. MS(El) m/e 565.9109 (4%, M⁺ =565.9091). ¹ HNMR (DMSO-d₆): δ 7.74 (2×d, 2, 6-H and 7-H, J₆₋₇ =8.5 Hz), 6.26 (d, 1,1'-H, J_(1'-2') =6.5 Hz), 5.53 (t, 1, 2'-H, J_(2'-3') =7.0 Hz), 5.42(dd, 1, 3'-H, J_(3'-4') =4.5 Hz), 4.42.

2-Chloro-4,5-dibromo-1-β-D-ribofuranosylbenzimidazole (85)

A solution of 0.529 g (0.930 mmol) of 84 in 25 mL of NH₃ /MeOH wasstirred in a pressure bottle at room temperature for 5 hr. Volatilematerials were removed by evaporation and coevaporation with MeOH (3×,bath temperature<40° C.). The resulting solid was recrystallized fromMeOH/H₂ O to give 0.316 g (2 crops, 77%) of 85 as white crystals. MP:167°-169° C. MS: (Cl) m/e 440.8837 (20%, MH⁺ =440.8852). ¹ H NMR(DMSO-d₆): δ 8.05 (d, 1, 7-H, J₇₋₆ =8.5 Hz), 7.59 (d, 1, 6-H), 5.89 (d,1, 1'-H, J_(1'-2') =7.5 Hz), 5.51 (d, 1, 2'-OH, J_(2'-2'OH) =6.5 Hz),5.28 (d, 1, 3'-OH, J_(3'-3'OH) =4.5 Hz), 5.27 (t, 1, 5'-OH, J_(5'-5'OH)=5.0 Hz), 4.40 (m, 1, 2'-H, J_(2'-3') =5.5 Hz), 4.13 (m, 1, 3'-H,J_(3'-4') =2.0 Hz), 4.00 (m, 1, 4'-H, J_(4'-5') =J_(4'-5") =3.5 Hz),3.68 (m, 2, 5'-H and 5"-H, J_(5'-5") =12.0 Hz). ¹³ H NMR (DMSO-d₆): δ141.70 (C2), 141.58 (C3a), 132.21 (C7a), 127.42 (C6), 118.06 (C5),114.08 (C4), 113.87 (C7), 89.45 (C1'), 86.34 (C4'), 71.74 (C2'), 69.62(C3'), 61.07 (C5'). Anal. Calcd. for C₁₂ H₁₁ Br₂ ClN₂ O₄ : C, 32.57; H,2.51; N, 6.33. Found: C, 32.70; H, 2.33; N, 6.33.

2-Chloro-5,6-diiodobenzimidazole (41)

Compound 6-amino-2-chloro-5-iodobenzimidazole (26a) (0.190 g, 0.647mmol) was dissolved in a mixture of conc. H₂ SO₄ /ice-H₂ O (2 mL/3 mL)at 0° C. To this mixture, was added dropwise a solution of NaNO₂ /H₂ O(0.134 g, 1.942 mmol/5 mL). The reaction mixture was stirred at roomtemperature for 1 hr. A solution of KI/H₂ O (0.537 g/5 mL) was addeddropwise and stirring was continued at room temperature for 3 h and then100° C. for 15 min. The reaction mixture was extracted with EtOAc (50mL×2). The EtOAc solution was washed with Na₂ S₂ O₃ /H₂ O (1 g/50 mL),sat. NaHCO₃ (50 mL), sat. NaCl solution (50 mL), dried (Na₂ SO₄), andevaporated. The residue was recrystallized from MeOH to give 0.169 g ofa yellowish crystalline compound. The mother liquor was evaporated andthe residue was chromatographed on a silica column (2×4 cm, elutedsuccessively with 1%, 2% MeOH/CHCl₃). Evaporation of fractions 4-6 (20mL per fraction) and recrystallization from MeOH gave an additional0.040 g of product. The total yield was 0.209 g (80%). MP: 228°-229° C.(dec). MS: (El) m/e 403.8064 (100%, M⁺ =403.8074). ¹ H NMR (DMSO-d₆): δ13.50 (Br s, 1, 1-NH) (s, 2, 4-H and 7-H). Anal. Calcd. for C₇ H₃ ClI₂N₂ : C, 20.79; H, 0.75; N, 6.93. Found: C, 20.73; H, 0.83; N, 6.74.

2-Chloro-5,6-diiodo-1-β-D-ribofuranosylbenzimidazole (107)

To a suspension of 0.230 g (0.569 mmol) of2-chloro-5,6-diiodobenzimidazole (41) in 5 mL of MeCN, was added 0.140mL (0.569 mmol) of BSA. The reaction mixture was stirred at 80° C. for15 min to give a clear solution. This solution was treated with 0.199 g(0.626 mmol) of 1,2,3,5-tetra-O-acetyl-β-D-ribofuranose and 0.132 mL(0.683 mmol) of TMSOTf at 80° C. for 45 min. The reaction mixture wascooled and diluted with EtOAc (50 mL). The EtOAc solution was washedwith sat. NaHCO₃ solution (50 mL×2), sat. NaCl solution (50 mL), dried(Na₂ SO₄), and evaporated. The residue was chromatographed on a silicacolumn (2.4×15 cm, eluted with CHCl₃). Evaporation of fractions 18-25(20 mL per fraction) and recrystallization from MeOH gave 0.157 g (42%)of the blocked nucleoside as white crystals. MP: 120°-123° C. MS: (El)m/e 661.8823 (14%, M⁺ =661.8814). ¹ H NMR (DMSO-d₆): δ 8.34 (s, 1, 7-H),8.25 (s, 1,4-H), 6.22 (d, 1, 1'-H, J_(1'-2') =7.0 Hz), 5.54 (t, 1, 2'-H,J_(2'-3') =7.0 Hz), 5.41 (dd, 1, 3'-H, J_(3'-4') =4.5 Hz), 4.46, 4.36(2×m, 3, 4'-H and 5'-H), 2.16, 2.14, 2.01 (3×s, 9, 3×Ac). ¹³ H NMR(DMSO-d₆): δ 169.94, 169.45, 169.15 (3×COCH₃), 142.54 (C3a), 140.67(C2), 134.08 (C7a), 128.88 (C4), 121.48 (C7), 101.98, 101.32 (C5 andC6), 86.53 (C1'), 79.48 (C4'), 70.51 (C2'), 68.71 (C3'), 62.57 (C5'),20.90, 20.27, 19.98 (3×COCH₃). Anal. Calcd. for C₁₈ H₁₇ ClI₂ N₂ O₇ : C,32.63; H, 2.59; N, 4.23. Found: C, 32.81; H, 2.63; N, 4.25. Thiscompound was deprotected to afford compound 107.

2,5,6-Trichloro-1-(2-deoxy-3,5-di-O-p-toluoyl-β-D-erythro-pentofuranosyl)benzimidazole(110)

To a suspension of 4.43 g (20 mmol) of 2,5,6-trichlorobenzimidazole (5)in 100 mL of dry MeCN, was add portionwise 1.2 g (30 mmol) of 60% NaH inoil at room temperature. After the addition had been completed, thereaction mixture was stirred at room temperature for 20 min. to give anearly clear yellowish solution. To this solution, compound3,5-di-O-p-toluyl-β-D-erythro-pentofuranosyl chloride (9.332 g, 24 mmol)was added portionwise over 20 min. and stirring was continued at roomtemperature for an additional 2 h. The reaction mixture was filtered andthe solid was washed with portions of EtOAc (˜300 mL). The filtrate wasevaporated and the residue was dissolved in the EtOAc washings. ThisEtOAc solution was washed with half sat. NaCl solution (150 mL×2), dried(Na₂ SO₄), and evaporated. The residue was added to 100 mL of MeOH, themixture was heated at reflux temperature for 5 min. and was then allowedto cool to room temperature. Filtration of the resulting suspension andwashing the solid with portions of MeOH gave 10.21 g (89%, 2 crops) of110 as white crystals. MP: 168°-169° C. MS: (El) m/e 572.0664 (0.5%, M⁺=572.0673). ¹ H NMR (DMSO-d₆): δ 8.04, 7.94 (2×s, 2, 7-H and 4-H), 7.97,7.86, 7.37, 7.29 (4×d, 8, Ph, J=8.0 Hz), 6.56 (dd, 1, 1'-H, J_(1'-2')=8.5 Hz, J_(1'-2") =6.0 Hz), 5.75 (m, 1, 3'-H, J_(3'-2') =8.0 Hz,J_(3'-2") =2.0 Hz, J_(3'-4') =3.5 Hz), 4.72 (dd, 1, 5'-H, J_(5'-4') =3.5Hz, J_(5'-5") =12.0 Hz), 4.69 (dd, 1, 5"-H, J_(5"-4') =5.0 Hz), 4.61 (m,1, 4'-H), 3.02 (m, 1, 2'-H, J_(2'-2") =14.5 Hz), 2.72 (m, 1, 2"-H),2.40, 2.36 (2×s, 6, 2×Me). ¹³ C NMR (DMSO-d₆): δ 165.48, 165.34(2×p-MePhCO), 144.05, 143.81 (2×p-MePhCO), 141.38 (C2), 140.81 (C3a),132.43 (C7a), 129.48, 129.24 (2×p-MePhCO), 126.47, 126.39, 126.23,125.91 (2×p-MePhCO, C6, and C5), 120.29 (C4), 113.43 (C7), 85.19 (C1'),80.07 (C4'), 73.52 (C3'), 63.72 (C5'), 35.75 (C2'), 21.14, 21.09(2×p-MePhCO). Anal. Calcd. for C₂₈ H₂₃ Cl₃ N₂ O₅ : C, 58.60; H, 4.04; N,4.88. Found: C, 58.35; H, 4.09; N, 4.63.

2,5,6-Trichlorobenzimidazole-1-2-deoxy-β-D-erythropentofuranosyl (111)

A suspension of 7.30 g (12.721 mmol) of 110 and 8.234 g (127.21 mmol) ofKCN in 255 mL of 90% aq. EtOH was stirred at room temperature for 4days. The reaction mixture was filtered and the filtrate was evaporated.The resulting solid was triturated successively with H₂ O (50 mL×3),hexane (50 mL×3), CHCl₃ (50 mL), and was then recrystallized from MeOHto give 3.027 g (70%, 2 crops) of 111 as white crystals. MP: 178°-180°C. MS: (El) m/e 335.9831 (12%, M⁺ =335.9835). ¹ H NMR (DMSO-d₆): δ 8.44(s, 1, 7-H), 7.94 (s, 1, 4-H), 6.35 (dd, 1, 1'-H, J_(1'-2') =9.0 Hz,J_(1'-2") =6.0 Hz), 5.42 (d, 1, 3'-OH, J_(3'-3'OH) =4.5 Hz), 5.24 (t, 1,5'-OH, J_(5'-5'OH) =5.0 Hz), 4.43 (m, 1, 3'-H, J_(3'-2') =7.0 Hz,J_(3'-2") =2.0 Hz, J_(3'-4') =2.5 Hz), 3.90 (m, 1, 4'-H, J_(4'-5') =3.0HZ), 3.70 (dd, 2, 5'-H), 2.51 (m, 1, 2'-H, J_(2'-2") =13.5 Hz), 2.19 (m,1, 2"-H). ¹³ C NMR (DMSO-d₆): δ 141.21 (C2), 140.95 (C3a), 132.27 (C7a),125.91, 125.67 (C5 and C6), 120.02 (C4), 114.77 (C7), 87.68 (C1'), 85.70(C1'), 69.99 (C3'), 60.86 (C5'), 38.96 (C2'). Anal. Calcd. for C₁₂ H₁₁Cl₃ N₂ O₃ : C, 42.69; H, 3.28; N, 8.30. Found: C, 42.40; H, 3.36; N,8.07.

2-Bromo-5,6-dichloro-1-(deoxy-β-D-erythro-pentofuranosyl)benzimidazole(112)

To a suspension of 1.55 g (5.829 mmol) of2-bromo-5,6-dichlorobenzimidazole (7) in 30 mL of dry MeCN, was addedportionwise 0.35 g (8.750 mmol) of 60% NaH in oil at room temperature.After the addition had been completed, the reaction mixture was stirredat room temperature for 20 min to give a nearly clear yellowishsolution. To this solution, the appropriate carbohydrate (2.72 g, 6.995mmol) was added portionwise over 20 min and stirring was continued atroom temperature for an additional 2.5 h. The reaction mixture wasdiluted with EtOAc (100 mL), filtered and the solid was washed withportions of EtOAc (20 mL). This EtOAc solution was washed with half sat.NaCl solution (100 mL×2), dried (Na₂ SO₄), and evaporated. The residuewas chromatographed on a silica column (4×16 cm, eluted with purechloroform). Evaporation of fractions 18-91 and recrystallization of theresidue from EtOH gave 2.927 g (81%, 2 crops) of the protectednucleoside as white crystals. MP: 157°-159° C. MS: (El) m/e 616.0153(0.2%, M⁺ =616.0167). ¹ H NMR (DMSO-d₆): δ 8.02, 7.95 (2×s, 2, 7-H and4-H), 7.96, 7.86, 7.37, 7.29 (4×d, 8, Ph, J=8.0 Hz), 6.52 (dd, 1, 1'-H,J_(1'-2') =9.0 Hz, J_(1'-2") =6.0 Hz), 5.76 (m, 1, 3'-H, J_(3'-2') =8.0Hz, J_(3'-2") =2.0 Hz, J_(3'-4') =3.5 Hz), 4.71 (m, 2, 5'-H and 5"-H,J_(5'-4') =4.0 Hz, J_(5"-4') =4.5 Hz, J_(5'-5") =12.0 Hz), 4.63 (m, 1,4'-H), 3.00 (m, 1, 2'-H, J_(2'-2") =14.5 Hz), 2.70 (m, 1, 2"-H), 2.40,2.36 (2×s, 6, 2×Me). ¹³ C NMR (DMSO-d₆): δ 165.45, 165.27 (2×p-MePhCO),144.00, 143.75 (2×p-MePhCO), 142.44 (C3a), 132.49 (C7a), 131.46 (C2),129.40, 129.19 (2×p-MePhCO), 126.43, 126.36, 126.03, 125.77 (2×p-MePhCO,C6, and C5), 120.12 (C4), 113.29 (C7), 65.27 (C1'), 80.97 (C4'), 73.50(C3'), 63.71 (C5'), 35.81 (C2') 21.08, 21.03 (2×p-MePhCO). Anal. Calcd.for C₂₈ H₂₃ BrCl₂ N₂ O₅ : C, 54.39; H, 3.75; N, 4.53. Found: C, 54.54;H, 3.59; N, 4.44.

A suspension of 0.618 g (1.0 mmol) of the protected nucleoside and 0.330g (5.0 mmol) of KCN in 20 mL of 90% aq. EtOH was stirred at roomtemperature for 5 days. The reaction mixture was evaporated. Theresulting solid was triturated successively with H₂ O (10 mL×3), CHCl₃(10 mL×3), and was then recrystallized from EtOH to give 0.300 g (79%, 3crops) of 112 as white crystals. MP: 187°-188° C. MS: (El) m/e 379.9332(6%, M⁺ =379.9330). ¹ H NMR (DMSO-d₆): δ 8.48 (s, 1,7-H), 7.93 (s,1,4-H), 6.33 (dd, 1, 1'-H, J_(1'-2') =9.0 Hz, J_(1'-2") =5.5 Hz), 5.48(d, 1, 3'-OH, J_(3'-3'OH) =4.0 Hz), 5.31 (t, 1, 5'-OH, J_(5'-5'OH) =4.5Hz), 4.43 (m, 1, 3'-H, J_(3'-2') =6.5 Hz, J_(3'-2") =1.5 Hz), 3.91 (m,1, 4'-H), 3.71 (m, 2, 5'-H), 2.50 (m, 1, 2'-H, J_(2'-2") =13.5 Hz), 2.15(m, 1, 2"-H). ¹³ C NMR (DMSO-d₆): δ 142.56 (C3a), 132.49 (C7a), 131.39(C2), 125.75, 125.56 (C5 and C6), 119.63 (C4), 114.66 (C7), 87.68 (C4'),86.94 (C1'), 70.02 (C3'), 60.86 (C5'), 39.00 (C2'). Anal. Calcd. for C₁₂H₁₁ BrCl₂ N₂ O₃ : C, 37.73; H, 2.90; N, 7.33. Found: C, 38.18; H, 2.80;N, 7.30.

5-Bromo-2,6-dichloro-1-β-D-ribofuranosylbenzimidazole (95)

To a suspension of 0.319 g (1.0 mmol) of2,6-dichloro-1-β-D-ribofuranosylbenzimidazole (67) in 10 mL of H₂ O, wasadded dropwise a sat. solution of Br₂ /H₂ O at room temperature. Afterthe addition had been completed, stirring was continued for 3 hr. Thereaction mixture was filtered and the solid was washed with portions ofH₂ O, and then recrystallized from MeOH to give 0.335 g (78%, as M MeOH)of 95 as white crystalline needles. MP 140°-142° C. MS (El) m/e 395.9274(2%, M⁺ =395.9279). ¹ H NMR (DMSO-d₆): δ 8.88 (s, 1, 7-H), 8.08 (s,1,4-H), 5.89 (d, 1, 1'-H, J_(1'-2') =8.0 Hz), 5.49 (d, 1, 2'-OH,J_(2'-2'OH) =6.5 Hz), 5.40 (t, 5'-OH, J_(5'-5'OH) =4.0 Hz), 5.28 (d, 1,3'-OH, J_(3'-3'OH) =4.5 HZ), 4.42 (m, 1, 2'-H, J_(2'-3') =5.5 Hz), 4.13(m, 1, 3'-H, J_(3'-4') =1.5 Hz), 4.01 (m, 1, 4'-H, J_(4'-5') =J_(4'-5")=2.5 Hz), 3.70 (m, 2, 5'-H and 5"-H, J_(5'-5") =12.0 Hz). Anal. Calcd.for C₁₂ H₁₁ BrCl₂ N₂ O₄ MeOH: C, 36.30; H, 3.51; N, 6.5 Found: C, 35.98;H, 3.60; N, 6.39.

6-Bromo-2,5-dichloro-1-β-D-ribofuranosylbenzimidazole (99)

To a suspension of 0.110 g (0.313 mmol, as C₁₂ H₁₂ Cl₂ N₂ O₄ MeOH) of2,5-dichloro-1-β-D-ribofuranosylbenzimidazole (73) in 3 mL of H₂ O wasadded dropwise 10 mL of a sat. solution of Br₂ /H₂ O at roomtemperature. After the addition had been completed, stirring wascontinued for 6 hr. The reaction mixture was filtered and the solid waswashed with portions of H₂ O, and then recrystallized from MeOH to give0.091 g (73%, 2 crops) of 99 as white crystalline needles. MP 158°-159°C. MS (El) m/e 395.9274 (5% M⁺ =395.9279). ¹ H NMR (DMSO-d₆): δ 8.69 (s,1, 7-H), 7.96 (s, 1,4-H), 5.88 (d, 1, 1'-H, J_(1'-2') =8.0 Hz), 5.51 (d,1, 2'-OH, J_(2'-2'OH) =6.5 Hz), 5.41 (t, 5'-OH, J_(5'-5'OH) =4.5 Hz),5.30 (d, 1, 3'-OH, J_(3'-3'OH) =4.5 Hz), 4.42 (m, 1, 2'-H, J_(2'-3')=5.5 Hz), 5.30 (d, 1, 3'-OH, J_(3'4') =1.5 Hz), 4.01 (m, 1, 4'-H,J_(4'-5') =J4'-5'=2.5 Hz), 371 (m, 2, 5'-H and 5"-H, J_(5'-5") =1.20Hz). Anal. Calcd. for C₁₂ H₁₁ BrCl₂ N₂ O₄ : C, 36.21; H, 2.79; N, 7.04.Found: C, 36.14; H, 2.90; N, 6.90.

4,5-Difluoro-1,2-phenylenediamine (10)

To a solution of 5.55 g (31.876 mmole) of 4,5-difluoro-2-nitroaniline in50 mL of MeOH, were added 100 mL of 2N HCl and 8.90 g (159.380 mmole) ofiron powder. The reaction mixture was stirred at room temperature for 2hr and then filtered. The filtrate was neutralized with conc. NH₄₀ Hto˜pH 8. The resulting suspension was filtered again and the filter cakewas washed thoroughly with MeOH. The filtrate and washings werecombined, concentrated to˜100 mL, and extracted with CHCl₃ (100 mL×3).The CHCl₃ solution was washed with a sat. NaCl solution (100 mL×2),dried (Na₂ SO₄), and evaporated. The residue was coevaporated with CHCl₃to give 3.515 g (77%) of 10. This material was used in the next reactionwithout further purification. A brown crystalline sample of 10 wasobtained by recrystallization from CCl₄. ¹ H NMR (DMSO-d₆): d 6.44 (t,2, 3-H and 6-H, J_(F-H) =10.5 H), 4.59 (br. s, 4, 2×NH₂).

2-Amino-5,6-difluorobenzimidazole (11)

To a stirred solution of 4.9 mL of 5M BrCN/MeCN in 55 mL of H₂ O, wasadded dropwise a solution of 3.515 g (24.389 mmole) of 10 in 50 mL ofMeOH over 20 min. After the addition, stirring was continued at roomtemperature for 2 h. The reaction mixture was concentrated to˜50 mL andwas extracted with EtOAc (50 mL×3). The combined EtOAc solution was backwashed with 100 mL of H₂ O and then discarded. The combined H₂ O phasewas basified with sat. NaHCO₃ solution (precipitation occurred) and wasextracted again with EtOAc (70 mL×3). The EtOAc solution was dried (Na₂SO₄) and evaporated to dryness, The residue was suspended in 50 mL ofCHCl₃ and filtered. The yellowish solid was washed with portions ofCHCl₃ to give 3.40 g of 11. The filtrate and washings were evaporated todryness and the procedure was repeated to give an additional 0.355 g of11. The total yield of 11 was 3.755 g (91%). ¹ H NMR (DMSO-d₆) d 10.79(br. s, 1, 1-NH), 7.06 (dd, 2, 4-H and 7-H, ³ J_(F-H) =11.0 Hz, ⁴J_(F-H) =7.5 Hz), 6.30 (br, s, 2, 2-NH₂).

2-Chloro-5,6-difluorobenzimidazole (12)

To a stirred mixture of 40 mL of aqueous CuCl₂ solution (60% by weight)and 20 mL of H₂ O was added a solution of NaNO₂ /H₂ O (2.08 g/10 mL).Compound 11 (1.69 g, 10 mmole) was then added portionwise over 5 min.The reaction mixture was stirred at room temperature for 1 hr. Anadditional 30 ml of the aqueous CuCl₂ solution (60% by weight) was addedand the reaction mixture was heated on a steam bath for 10 min (a smallamount of MeOH was added to suppress the formation of foam). The mixturewas extracted with EtOAc (150 mL×2). The EtOAc solution was washed withsat. NaCl solution (100 mL×2), dried (Na₂ SO₄) and evaporated. Theresidue was chromatographed on a silica column (3×15 cm) using 2% and 3%MeOH/CHCl₃ as eluants. Evaporation of the appropriate fractions gave abrownish solid. This solid was washed with Et₂ O and dried to give 0.962g of 12. The Et₂ O washings were evaporated and the residue wasrepurified through a silica column (2×10 cm, eluted successively with1%, 2% MeOH/CHCl₃). Evaporation of the appropriate fractions and washingof the residue with Et₂ O gave an additional 0.152 g of 12. The totalyield of 12 was 1.114 g (59%). MS m/e 187.9954 (100%, M⁺ =187.9953). ¹ HNMR (DMSO-d₆): d 13.5 (br. s, 1, 1-NH), 7.62 (t, 2, 4-H and 7-H, J_(F-H)=9.0 Hz.

2-Amino-5(6)-chloro-6(5)-fluorobenzimidazole (12b)

4-Chloro-5-fluoro-1,2-phenylenediamine (12a, 3.212 g, 20 mmol) wasdissolved in 40 mL of MeOH and then added dropwise to a stirred solutionof 4.4 mL of 5M BrCN/MeCN in 40 mL of H₂ O over 30 min. After theaddition, stirring was continued at room temperature for 3 hr. Thereaction mixture was concentrated to˜40 mL and then was washed withEtOAc (100 mL). The EtOAc phase was extracted with H₂ O (60 mL). Thecombined H₂ O phase was neutralized with sat. NaHCO₃ solution to˜pH 8and the resulting suspension was extracted with EtOAc (200 mL). TheEtOAc phase was washed with a mixture of sat. NaHCO₃ /sat. NaCl solution(20 mL/180 mL), dried (Na₂ SO₄), and evaporated. The residue wascoevaporated with CHCl₃ (213 mL×2) and then was suspended in 50 mL ofCHCl₃. The suspension was filtered to give 3.340 g of 12b as a greysolid. The filtrate was evaporated and coevaporated with CHCl₃. Theresulting solid was again suspended in a small amount of CHCl₃ and thesuspension was filtered to give an additional 0.235 g of 12b. The totalyield of 12b was 3.575 g (96%). This product showed a single spot onTLC. MP: 194°-196° C. MS: (El) m/e 185.0156 (100%, M⁺ =185.0156). ¹ HNMR (DMSO-d₆): d 10.85, (br s, 1, 1-NH), 7.16 (d, 1,4-H, ⁴ J_(F-H) =7.0Hz), 7.06 (d, 1,7-H, ³ J_(F-H) =10.0 Hz), 6.43 (br s, 2, 2-NH₂). Anal.Calcd. for C₇ H₅ ClFN₃ : C, 45.30; H, 2.72; N, 22.64. Found: C, 45.14;H, 2.67; N, 22.44.

2,5(6)-Dichloro-6(5)-fluorobenzimidazole (12c)

Compound 12b (0.928 g, 5.0 mmol) was suspended in a solution of CuCl₂/H₂ O (10 g/25 mL). To this suspension, was added dropwise a solution ofNaNO₂ /H₂ O (3.45 g, 50 mmol/25 mL) at 70° C. over 30 min (5 mL oft-BuOH was added portionwise to suppress foaming). After the addition,stirring was continued at 70° C. for 15 min. The reaction mixture wascooled and extracted with EtOAc (75 mL×2). The EtOAc solution wasfiltered and the filtrate was washed with half sat. NaCl solution (100mL), dried (Na₂ SO₄), and evaporated. The residue was chromatographed ona silica column (2.4×25 cm, eluted successively with 1%, 2% MeOH/CHCl₃).Evaporation of fractions 23-35 (20 mL per fraction) gave 0.744 g (73%)of 12c as a yellowish solid. MP: 197°-198° C. MS: (El) m/e 203.9657(100%, M⁺ =203.9657). ¹ H NMR (DMSO-d₆): d 13.60 (br s, 1, 1-NH), 7.76(d, 1,4-H, ⁴ J_(F-H) =7.0 Hz), 7.61 (d, 1,7-H, ³ J_(F-H) =10.5 Hz).Anal. Calcd. for C₇ H₃ Cl₂ FN₂ : C, 41.01; H, 1.47; N, 13.66. Found: C,41.00; H, 1.39; N, 13.59.

2,4,5,6-Tetrachlorobenzimidazole (13)

To a mixture of CuCl₂ (1.345 g, 10 mmol) and 0.99 mL of 90% t-BuONO (30mmol) in 25 mL of acetone, was added portionwise 1.280 g (5 mmol) of 13bover a period of 3 min. After the addition had been completed, stirringwas continued at 60° C. for 2 h (with the addition of 0.99 mL of fresh90% t-BuONO every 30 min). The reaction mixture was cooled to roomtemperature, poured into 50 mL of 2N HCl, and extracted with 150 mL ofEtOAc. The EtOAc layer was washed with 2N HCl (50 mL×2), sat. NaClsolution (100 mL×2), dried (Na₂ SO₄), and evaporated. The residue waschromatographed on a silica column (4×7 cm, eluted with CHCl₃, 1%MeOH/CHCl₃). Evaporation of fractions 31-45 (15 mL per fraction) andrecrystallization from MeOH gave 0.631 g (2 crops, 49%) of 13 asslightly brownish crystals. MP: 243°-244° C. MS: (El) role 253.8952(81%, M⁺ =253.8972). ¹ H NMR (DMSO-d₆): d 14.09 (br s, 1, 1-NH), 7.83{s, 1, 7(4)-H}. Anal. Calcd. for C₇ H₂ Cl₄ N₂ : C 32.85, H 0.78, N10.95. Found: C, 32.69; H, 0.84; N, 10.76.

2-Amino-4,5,6-trichlorobenzimidazole (13b)

To a stirred solution of 3.62 mL of 5M BrCN/MeCN in 35 mL of H₂ O, wasadded dropwise a solution of 3,4,5-trichloro-1,2-phenylenediamine (13a)(3.478 g, 16.446 mmol) in 35 mL of MeOH. After the addition had beencompleted, stirring was continued at room temperature for 2 hr. Thereaction mixture was concentrated to .sup.˜ 35 mL and was washed withEtOAc (50 mL). The EtOAc phase was extracted with H₂ O (50 mL). Thecombined H₂ O phase was neutralized with sat. NaHCO₃ solution to .sup.˜pH 8 and the resulting suspension was extracted with EtOAc (150 mL). TheEtOAc phase was washed with a mixture of sat. NaHCO₃ and sat. NaClsolution (213 mL/130 mL), dried (Na₂ SO₄), and evaporated. The residuewas dissolved in MeOH, decolorized with charcoal, and thenrecrystallized from MeCN to give 3.098 g (2 crops, 80%) of 13b as aslightly grey crystals. MP: 255°-258° C. MS: (El) m/e 234.9473 (100%, M⁺=234.9471). ¹ H NMR(DMSO-d₆): d 11.25 (br s, 1, 1-NH), 7.29 {s, 1,7(4)-H}, 6.75 (br s, 2, 2-NH₂). Anal. Calcd. for C₇ H₄ Cl₃ N₃ : C 35.55,H 1.70, N 17.77, Found: C 35.72, H 1.78, N 17.89.

2-Chloro-5,6-dibromobenzimidazole (17)

To a suspension of 0.763 g (5 mmole) of 2-chlorobenzimidazole in 50 mLof 1:1 MeOH/H₂ O, was added dropwise a solution of 1 mL of Br₂ in 10 mLof MeOH over a period of 30 min. The reaction mixture was stirred atroom temperature overnight and was then filtered. The solid was washedwith portions of H₂ O until the washings were neutral. This solid wasair dried and recrystallized from MeOH to give 1.115 g (3 crops, 72%) of17. mp 228° C.; ¹ H NMR (DMSO-d₆) d 13-14 (br. s, 1, 1-NH), 7.93 (s, 2,4-H, 7-H).

2-Chloro-5-nitrobenzimidazole (19)

To 210 mL of fuming HNO₃, 2-chlorobenzimidazole (13.42 g, 87.95 mmole)was added portionwise over a period of 10 min with stirring and ice-H₂ Ocooling. After the addition, the cooling bath was removed and stirringwas continued at room temperature overnight. This reaction mixture wascooled to .sup.˜ 0° C., poured into 300 mL of ice, and neutralizedcarefully with conc. NH₄ OH to .sup.˜ pH 8. The resulting suspension wasfiltered. The yellowish solid product was washed with portions of H₂ Oand air-dried. The filtrate and washings were combined and extractedwith EtOAc (200 mL×2). The EtOAc solution was washed with sat. NaClsolution (200 mL×2), dried (Na₂ SO₄), and evaporated to give 1.3 g of ayellowish solid. The solid was combined with the major part of theproduct and was recrystallized from MeOH to give 16.34 g (5 crops, 94%)of 19 as yellowish crystals. mp 235°-237° C. MS (El) m/e 196.9984 (100%,M⁺ =196.9992). ¹ H NMR (DMSO-d₆) d 14.06 (br. s, 1, 1-NH), 8.41 (s, 1,4-H), 8.14 (dd, 1, 6-H, J₆₋₄ =2.0 Hz, J₆₋₇ =9.0 Hz), 7.71 (d, 1, 7-H).

2-Chloro-4,5,6-tribromobenzimidazole (20)

To a suspension of 4.578 g (30 mmol) of 2-chlorobenzimidazole (16) in amixture of MeOH/t-BuOH/H₂ O (20 mL/25 mL/100 mL), was added dropwise asolution of Br₂ /MeOH (7.728 mL/20 mL) over 2 h. After the addition hadbeen completed, the reaction mixture was Stirred at room temperature forI day. More solvents (MeOH/H₂ O, 25 mL/100 mL) were added, stirring wascontinued at room temperature for 2 days. Fresh Br₂ /MeOH (1.546 mL/10mL) was added, stirring was continued at room temperature for 1 day andthen at 50° C. for 6 hr. The reaction mixture was cooled and filtered..The filter cake was washed with portions of H₂ O, air-dried, andfractionally recrystallized from MeOH to give 1.70 g (15%) of 20 ascrystalline needles (mainly one spot on TLC). {the major fraction (6.154g) was a mixture of compounds 17, 20, and2-chloro-4,5,6,7-tetrabromobenzimidazole (23}. MP: 263°-266° C. ¹ H NMR(DMSO-d₆): d 14.00 (br s, 1, 1-NH), 7.96 {s, 1, 7(4)-H}.

5(6)-Amino-2-chloro-6(5)-nitrobenzimidazole (25)

A mixture of 1.213 g (5.0 mmol) of 2-chloro-5,6-dinitrobenzimidazole(22) and 1.398 g (25 mmol) of iron powder in 50 mL of AcOH was stirredat room temperature for 4 h. The reaction mixture was diluted with 100mL of EtOAc, filtered, and the solid was washed with portions of EtOAc(total EtOAc used .sup.˜ 100 mL). The filtrate and washings werecombined and washed with H₂ O (100 mL×3). The H₂ O layer was extractedwith 100 mL of EtOAc. The EtOAc solutions were combined, evaporated,coevaporated with toluene (10 mL×3), MeOH (10 mL×2) to give a brownsolid.

The brown solid was absorbed on 30 mL of silica gel and waschromatographed on a silica column (3×25 cm, eluted successively with2%, 4%, 8% MeOH/CHCl₃). Evaporation of fractions 30-58 (20 mL perfraction) and recrystailization from MeOH gave 0.431 g (2 crops, 41%) of25 as red crystals. MP: >250° C. (dec). MS: (El) m/e 212.0096 (100%, M⁺=212.0101). ¹ H NMR (DMSO-d₆): d 13.07 (br s, 1, 1-NH), 8.15 (s, 1,7-H),7.06 (s, 2,5-NH₂), 6.91 (s, 1,4-H). Anal. Calcd. for C₇ H₅ ClN₄ O₂ : C,39.55; H, 2.37; N, 26.35. Found: C, 39.81; H, 2.11; N, 26.43.

2-Chloro-5(6)-iodo-6(5)-nitrobenzimidazole (26)from2-chloro-5,6-dinitrobenzimidazole (22)

A mixture of 1.213 g (5.0 mmol) of 22 and 1.398 g (25 mmol) of ironpowder in 50 mL of AcOH was stirred at room temperature for 4 hr. Thereaction mixture was diluted with 100 mL of EtOAc, filtered, and thefiltrate was washed with portions of EtOAc (total EtOAc used .sup.˜ 100mL). The filtrate and washings were combined and washed with H₂ O (50mL×3). The H₂ O layer was extracted with 100 mL of EtOAc. The EtOAcsolutions were combined, evaporated, coevaporated with toluene (10mL×3), MeOH (10 mL×2) to give a brown solid.

The brown solid was dissolved in a mixture of Conc. H₂ SO₄ /ice-H₂ O (14mL/20 mL) at 0° C. To this mixture, was added dropwise a solution ofNaNO₂ /H₂ O (0.994 g, 13.688 mmol/5 mL). The reaction mixture wasstirred at 0° C. for 1 hr. A solution of urea/H₂ O (0.411 g/3 mL) wasadded and stirring was continued at 0° C. for 10 min. A solution ofKl/H₂ O (2.272 g/5 mL) was added and stirring was continued at roomtemperature for 18 h. The reaction mixture was extracted with EtOAc (100mL×2). The EtOAc solution was washed with H₂ O (100 mL), sat. NaHCO₃(100 mL), sat. NaCl solution (100 mL), dried (Na₂ SO₄), and evaporated.The residue was chromatographed on a silica column (2.2×25 cm, elutedsuccessively with 1%, 2% MeOH/CHCl₃). Fractions 20-41 (20 mL perfraction) were collected, washed with Na₂ S₂ O₃ /H₂ O (1 g/100 mL),dried(Na₂ SO₄), and evaporated. The residue was recrystallized from MeOHto gave 0.593 g (3 crops, 40%) of 26 as yellowish crystals. MP: .sup.˜213° C. (dec). MS: (El) role 322.8968 (100%, M⁺ =322.8959). ¹ H NMR(DMSO-d₆): d 13.98 (br s, 1, 1-NH), 8.24, 8.16 (2 x s, 2,4-H and 7-H).Anal. Calcd. for C₇ H₃ CllN₃ O₂ : C, 25.99; H, 0.93; N, 12.99. Found: C,25.74; H, 0.73; N, 12.71.

6(5)-Amino-2-chloro-5(6)-iodobenzimidazole (26a)

A mixture of 0.339 g (1.048 mmol) of 26 and 0.090 g (wet) of Ra--Ni in10 mL of EtOH was hydrogenated at room temperature, 50 psi of H₂ for 7h. The reaction mixture was filtered and the filtrate was evaporated.The residue was chromatographed on a silica column (2×6 cm, elutedsuccessively with 1%, 2%, 3% MeOH/CHCl₃). Evaporation of fractions 11-17(20 mL per fraction) gave 0.190 g (62%) of 26a as a foam. An analyticalsample (a yellowish crystalline compound) was obtained byrecrystallization from MeOH. MP: >200° C. (dec). MS: (El) m/e 292.9202(100%, M⁺ =292.9217). ¹ H NMR (DMSO-d₆): d 12.80 (br s, 1, 1-NH), 7.75(s, 1, 4-H), 6.87 (s, 1, 7-H), 5.03 (s, 2, 6-NH₂). Anal. Calcd. for C₇H₅ CllN₃ : C, 28.65; H, 1.72; N, 14.32. Found: C, 28.84; H, 1.66; N,14.17.

2,4,6-Trichlorobenzimidazole (32)

To a solution of CuCl₂ /H₂ O (45 g/100 mL), were added a solution ofNaNO₂ /H₂ O (4.14 g, 60 mmol/20 mL) and then a solution of 29/MeOH (4.04g, 20 mmol/20 mL). After the addition, stirring was continued at roomtemperature for 1 hr. The reaction mixture was then heated on a steambath for 1 hr. During this period of heating, an additional fresh NaNO₂/H₂ O solution (4.14 g, 60 mmol/20 mL) was added dropwise. The reactionmixture was cooled and extracted with EtOAc (200 mL×2). The EtOAcsolution was filtered and the filtrate was washed with half sat. NaClsolution (200 mL×2), dried (Na₂ SO₄), and evaporated. The residue waschromatographed on a silica column (4×25 cm, eluted with 2% MeOH/CHCl₃).Evaporation of fractions 46-68 (20 mL per fraction) gave 1.796 g (41%)of 8 as a white foam. An analytical sample was obtained byrecrystallization from MeOH. MP: 233°-234° C. MS: (El) m/e 219.9357(100%, M⁺ =219.9362). ¹ H NMR (DMSO-d₆): d 13.92 (br s, 1, 1-NH), 7.58("s", 1, 5-H or 7-H), 7.44 ("d", 1, 5-H or 7-H, J₅₋₇ =1.5 Hz). Anal.calcd. for C₇ H₃ Cl₃ N₂ : C 37.96, H 1.37, N 12.65. Found: C 38.13, H1.68, N 12.67.

2-Amino-5(6)-chlorobenzimidazole (34)

To a solution of 5M BrCN/CH₃ CN (10 ml) in 100 ml of H₂ O) was addeddropwise a solution of 7.13 grams (50 mmoles) of4-chloro-o-phenylenediamine in methanol (100 ml) over a period of 30minutes while stirring. The contents were reacted at room temperaturefor an additional two hours. The reaction solution was concentrated toapproximately 100 ml and extracted with EtOAc (200 ml/100 ml H₂ O). Theorganic phase was extracted a second time with an additional 100 ml ofdistilled water and then discarded. The combined water phases wereneutralized with 40 ml of saturated sodium bicarbonate. The resultantprecipitate was collected by filtration. The filter cake was washed withportions of H₂ O (100 ml) and air dried. The filtrate and washings werecombined and extracted with EtOAc (100 ml×2). The EtOAc solution wasdried (Na₂ SO₄) and evaporated to give a solid. This was combined withthe major part of the solid product and was coevaporated with EtOH (2×).The resulting solid was suspended in 100 ml of CHCl₃ and the suspensionwas filtered. The filter cake was washed with portions of CHCl₃, and airdried to give 6.62 grams of 34 as a yellowish solid. The filtrate andwashings were evaporated and the residue was again suspended in a smallmount of CHCl₃. Filtration of the suspension gave an additional 0.88grams of 34 as a yellowish solid. This material was used for subsequentreactions without further purification. The total yield of 34 was 7.50grams (90%). mp 159°-161° C. MS (El) role 167.0243 (100%, M⁺ =167.0250).¹ H NMR (DMSO-d₆) d 10.81 (br. s, 1, 1-NH), 7.09 (d, 1, 4-H, J₄₋₆ =2.0Hz), 7.06 (d, 1,7-H, J₇₋₆ =8.5 Hz), 6.84 ("d", 1, 6-H), 6.33 hr. s, 2,2-NH₂).

2,5(6)-Dichlorobenzimidazole (35)

2-Amino-5(6)-chlorobenzimidazole (34) (10.06 grams, 60 mmoles) was addedportionwise to an aqueous solution of cupric chloride (120 grams) andsodium nitrite (12.42 grams) while stirring. The reaction mixture wasstirred for one hour at room temperature. At this time the reactionmixture was extracted with EtOAc (200 ml×3) to provide a brownishdiscoloration of the organic phase. The EtOAc phase was washed withsaturated NaCl solution, dried with Na₂ SO₄, and evaporated in vacuo toreveal a brown precipitate which was added to a silica column. Elutionwith 2% MeOH/CHCl₃, while increasing the polarity gradually to 5%MeOH/CHCl₃, and evaporation of the appropriate fractions provided abrownish-white precipitate which was suspended in Et₂ O and filtered toyield 53.1% (grams) of 35. mp 209°-212° C. MS (El) m/e 185.9751 (100%,M⁺ =185.9752). ¹ H NMR (DMSO-d₆) d 13.5 (br. s, 1, 1-NH), 7.59 (d, 1,4-H, J₄₋₆ =2.0 Hz), 7.52 (d, 1, 7-H, J₇₋₆ =8.5 Hz), 7.25 (m, 1, 6-H).

2-Chloro-5,6-diiodobenzimidazole (41)

Compound 26a (0.190 g, 0.647 mmol) was dissolved in a mixture of conc.H₂ SO₄ /ice-H₂ O (2 mL/3 mL) at 0° C. To this mixture, was addeddropwise a solution of NaNO₂ /H₂ O (0.134 g, 1.942 mmol/5 mL). Thereaction mixture was stirred at room. temperature for 1 hr. A solutionof Kl/H₂ O (0.537 g/5 mL) was added dropwise and stirring was continuedat room temperature for 3 h and then 100° C. for 15 min. The reactionmixture was extracted with EtOAc (50 mL×2). The EtOAc solution waswashed with Na₂ S₂ O₃ /H₂ O (1 g/50 mL), sat. NaHCO₃ (50 mL), sat. NaClsolution (50 mL), dried (Na₂ SO₄), and evaporated. The residue wasrecrystallized from MeOH to give 0.169 g of 41 as a yellowishcrystalline compound. The mother liquor was evaporated and the residuewas chromatographed on a silica column (2×4 cm, eluted successively with1%, 2% MeOH/CHCl₃). Evaporation of fractions 4-6 (20 mL per fraction)and recrystallization from MeOH gave additional 0.040 g of 41. The totalyield of 41 was 0.209 g (80%). MP: 228°-229° C. (dec). MS: (El) m/e403.8064 (100%, M⁺ =403.8074). ¹ H NMR (DMSO-d₆): d 13.50 (br s, 1, 1-NH), 8.11 (s, 2, 4-H and 7-H). Anal. Calcd. for C₇ H₃ Cll₂ N₂ : C,20.79; H, 0.75; N, 6.93. Found: C, 20.73; H, 0.83; N, 6.74.

2-Amino-4(7)-chloro-6(5)-trifluoromethylbenzimidazole (41b)

To a stirred solution of 4.8 mL of 5M BrCN/MeCN in 40 mL of H₂ O, wasadded dropwise a solution of3-chloro-5-trifluoromethyl-1,2-phenylenediamine (41a) (4.212 g, 20 mmol)in 40 mL of MeOH at room temperature over 30 min. After the addition hadfinished, stirring was continued at room temperature for 3 hr. Thereaction mixture was concentrated to .sup.˜ 40 mL and then was washedwith EtOAc (50 mL). The EtOAc phase was extracted with H₂ O (50 mL). Thecombined H₂ O phase was neutralized with sat. NaHCO₃ solution to .sup.˜pH 8 and the resulting suspension was extracted with EtOAc (100 mL×2).The EtOAc phase was washed with sat. NaCl solution (150 mL), dried (Na₂SO₄), and evaporated to give 3.913 g (83%) of 41b as a white foam. Thisproduct showed a single spot on TLC. An analytical sample was obtainedby addition of CHCl₃ to the above foam to effect crystallization. MP:180°-182° C. MS: (El) m/e 235.0121 (100%, M⁺ =235.0124). ¹ H NMR(DMSO-d₆): d 11.77, 11.25 (2×br s*, 1, 1-NH), 7.35, 7.26 (2×s, 2, Ph),6.90, 6.53 (2×br s*, 2, 2-NH₂). Anal. Calcd. for C₈ H₅ ClF₃ N₃ : C,40.79; H, 2.14; N, 17.84. Found: C, 40.90; H, 2.34; N, 17.79.

2,4(7)-Dichloro-6(5)-trifluoromethylbenzimidazole (41c)

To a solution of CuCl₂ /H₂ O (20 g/50 mL), was added a solution of NaNO₂/ H₂ O (3.45 g, 50 mmol/20 mL). Compound 41b (2.356 g, 10 mmol) was thenadded portionwise over 10 min (2 mL of t-BuOH was added to help themix). After the addition had finished, stirring was continued at roomtemperature for 1 h. The reaction mixture was then heated at 70° C. for30 min. During this period of heating, an additional fresh NaNO₂ /H₂ Osolution (3.45 g, 50 mmol/20 mL) was added dropwise. The reactionmixture was cooled and extracted with EtOAc (75 mL×2). The EtOAcsolution was filtered and the filtrate was washed with sat. NaClsolution (100 mL×2), dried (Na₂ SO₄), and evaporated. The residue waschromatographed on a silica column (3×25 cm, eluted with CHCl₃, 1%MeOH/CHCl₃). Evaporation of fractions 35-42 (20 mL per fraction) gave1.369 g (54%) of 41c as a yellowish solid. An analytical sample wasobtained by recrystallization from MeOH. MP: 187°-189° C. MS: (El) role253.9629 (100%, M⁺ =253.9625). ¹ H NMR (DMSO-d₆): d 14.22 (br s, 1,1-NH), 7.86, 7.68 (2×s, 2, Ph). Anal. Calcd. for C₈ H₃ Cl₂ F₃ N₂ : C,37.68; H, 1.19; N, 10.98. Found: C, 37.81; H, 1.08; N, 11.06.

1-(5-O-Acetyl-β-D-ribofuranosyl)-2,5,6-trichlorobenzimidazole (42a)

A mixture of 42 (0.48 g, 1.0 mmol) and KOAc (0.49 g; 5.0 mmol) inMeOH/H₂ O (20 mL/2 mL) was stirred at room temperature for 24 h. AcOH(0.286 mL, 5.0 mmol) was added and stirring was Continued at roomtemperature for 15 min. The reaction mixture was evaporated and theresidue was partitioned between H₂ O/CHCl₃ (50 mL/50 mL). The CHCl₃layer was washed with sat. NaCl solution (50 mL), dried (Na₂ SO₄), andevaporated. The residue was chromatographed on a silica column (1.9×20cm, eluted with CHCl₃, 1%, 2%, 4% MeOH/CHCl₃). Evaporation of fractions42-51 (15 mL per fraction) and recrystallization from MeOH gave 0.178 g(2 crops, 45%) of 42a as white crystals. MP: 82°-87° C. MS: (El) m/e393.9875 (20%, M⁺ =393.9890). ¹ H NMR (DMSO-d₆): d 8.00, 7.96 (2×s, 2,7-H and 4-H), 5.91 (d, 1, 1-H, J_(1'-2') =7.5 Hz), 5.59 (d, 1,2'-OH,J_(2'-2'OH) =6.0 Hz), 5.45 (d, 1, 3'-OH, J_(3'-3'OH) =4.5 Hz), 4.44 (dd,1, 5'-H, J_(5'-4') =4.5 Hz, J_(5'-5') =12.5 Hz), 4.42 (m, 1,2'-H,J_(2'-3') =6.0 Hz), 4.25 (dd, 1,5"-H, J_(5"-4') =2.5 Hz), 4.17 (m, 1,4'-H, J_(3'-4') =3.5 Hz), 4,11 (m, 1, 3'-H), 2.14 (s, 3, 5'-OAc) ¹³ CNMR (DMSO-d₆): d 169.95 (COCH₃), 142.03 (C2), 140.92 (C3a), 132.30(C7a), 125.96, 125.86 (C5 and C6), 120.28 (C4), 113.47 (C7), 89.32(C1'), 82.63 (C4'), 71.41 (C2'), 69.03 (C3'), 63.35 (C5'). 20.56(COCH₃). Anal. Calcd. for C₁₄ H₁₃ Cl₃ N₂ O₅ : C, 42.50; H, 3.31; N,7.08. Found: C, 42.45; H, 3.20; N, 6.97.

2-Amino-5,6-dichloro-1-(2,3,5-tri-O-acetyl-β-D-ribofuranosyl)benzimidazole (43)

2-Amino-5,6-dichlorobenzimidazole (4) (3 g, 16 mmole) was dissolved indry acetonitrile (150 ml) and stirred in an inert atmosphere at 60° C.BSA (4.37 ml, 17 mmole) was added and the mixture was stirred for 10minutes. 1,2,3,5-tetra-O-acetyl-β-D-ribofuranose (5.09 g, 16 mmole) andTMSTf (3.29 ml, 17 mmole) were added to the clear solution and themixture was allowed to stir at 60° C. for 1 hr. The mixture wasconcentrated under reduced pressure and separated on a silica column toyield 1.14 g (15%) of2-amino-5,6-dichloro-1-(2,3,5,tri-O-acetyl-β-D-ribofuranosyl)benzimidazole (43). ¹³ C NMR (CDCl₃): d 170.04 ppm, 169.56, 169.11,154.57, 141.64, 132.53, 125.90, 1123.45, 117.72, 109.49, 85.78, 80.99,70.95, 69.83, 62.91, 20.74, 20.54, 20.20. ¹ H NMR (CDCl₃): d 0.96 ppm(s, 3H), 1.09 (s, 3H), 1.13 (s, 3H), 3.75 (m, 2H), 4.08 (dd, 1H), 5.00(dd, 1H), 5.13 (t, 1H), 5.42 (s, 2H), 5.61 (d, 1H), 7.27 (s, 1H), 7.42(s, 1H). MS (FAB): m/e 758, 718, 676, 460, 426, 259, 217, 139.

2-Amino-5,6-dichloro-1-(β-D-ribofuranosyl)benzimidazole (44)

2,5,6-Trichloro-1-(2,3,5-tri-O-acetyl-β-D-ribofuranosyl)benzimidazole(42, 0.48 g; I mmole) was dissolved in dry MeOH (10 ml). Liq. NH₃ (10ml) was added and the mixture heated in a steel bomb at 100° C. for 2hr. This mixture was stirred at room temperature for 24 hr. Excess NH₃was allowed to evaporate and the solution evaporated to dryness,absorbed onto silica gel (2 g), and chromatographed on a silica gelcolumn using silica gel (20 g; 60-200 mesh). Elution of the column withCH₂ Cl₂ :CH₃ OH (96:4) gave the desired compound 44 as the majornucleoside. Evaporation of the solvent and crystallization of theresidue from EtOH gave 44 (0.02 g; yield 6.06%). mp. 145° C.; UVI_(max). (MeOH): 259 (7941) and 301 (8348) nm; I_(max) (pH1): 215(26000) and 292 (7520) nm; I_(max) (pH11): 257 (7967) and 298 (7107) nm.¹ H NMR(DMSO-d₆): d 8.70 (s,1H, C₇ -H, 8.28(s,1H, C₄ -H), 7.90 (bs,2H,D₂ O exchangeable, NH₂), 6.75(d,C1'H, J₁,2 =4.5 H_(z)) and the rest ofthe sugar protons.

1-(5-O-Acetyl-β-D-ribofuranosyl)-2-bromo-5,6-dichlorobenzimidazole (52b)

A mixture of 52a (1.048 g, 2.0 mmol) and KOAc (0.98 g, 10.0 mmol) inMeOH/H₂ O (40 mL/4 mL) was stirred at room temperature for 24 hr. AcOH(0.572 mL, 10.0 mmol) was added and stirring was continued at roomtemperature for 15 min. The reaction mixture was evaporated and theresidue was partitioned between. EtOAc/H₂ O (75 mL/75 mL). The EtOAclayer was washed with sat. NaCl solution (75 mL), dried (Na₂ SO₄), andevaporated. The residue was chromatographed on a silica column (1.9×24cm, eluted with CHCl₃, 2%, 3%, 4% MeOH/CHCl₃). Evaporation of fractions15-27 (15 mL per fraction) and recrystallization from MeOH gave 0.50 g(2 crops, 57%) of 52b as white crystals. MP: 85°-95° C. (melted slowlyover a large range) MS: (El) m/e 437.9398 (15%, M⁺ =437.9385). ¹ H NMR(DMSO-d₆): d 8.00, 7.95 (2×s, 2, 7-H and 4-H), 5.90 (d, 1, 1'-H,J_(1'-2') =7.5 Hz), 5.58 (d, 1,2'-OH, J_(2'-2') OH=6.5 Hz), 5.46 (d, 1,3'-OH, J_(3'-3'OH) =4.5 Hz), 4.44 (dd, 1, 5'-H, J_(5'-4') =4.5 Hz,J_(5'-5') =12.5 Hz), 4.43 (m, 1,2'-H, J_(2'-3') =6.0 Hz), 4.24 (dd,1,5"-H, J_(5"-4') =2.0 Hz), 4.16 (m, 1, 4'-H, J_(3'-4') =3.0 Hz), 4.11(m, 1, 3'-H), 2.15 (s, 3, 5'-OAc). ¹³ C NMR (DMSO-d₆): d 169.96 (COCH₃),142.57 (C3a), 132.49, 132.42 (C2 and C7a), 125.78, 125.74 (C5 and C6),120.14 (C4), 113.28 (C7), 90.32 (C1'), 82.56 (C4'), 71.27 (C2'), 68.99(C3'), 63.34 (C5'), 20.58 (COCH₃). Anal. Calcd. for C₁₄ H₁₃ BrCl₂ N₂ O₅: C, 38.21; H, 2.98; N, 6.37. Found: C, 38.37; H, 2.86; N, 6.27.

5,6-Dichloro-1-(β-D-ribofuranosyl)benzimidazol-2-thione (53)

A mixture of dry 5,6-dichlorobenzimidazole-2-thione (3, 2.19 g 10mmole), hexamethyldisilazane (4 ml) and (NH₄)₂ SO₄ (0.1 g) was heated atreflux temperature with stirring for 15 hr under anhydrous conditions.The clear brown reaction mixture was fractionated to remove unreactedHMDS under reduced pressure (water aspirator) to afford the disilylatedcompound. The silylated compound was mixed with1-bromo-2,3,5-tri-O-acetyI-D-ribofuranose (from 3.5 g (11 mmole) oftetraacetylsugar) and sodium iodide (0.05 g). The mixture was then fusedat 110° C. (oil bath temperature) with stirring for 45 minutes underreduced pressure. The reaction mixture which had been cooled to roomtemperature was dissolved in CHCl₃. The CHCl₃ solution was washed with acold saturated aqueous sodium bicarbonate solution (100 ml×4) and thencold water (100 ml×4). The CHCl₃ phase was dried over Na₂ SO₄ andevaporated on a water bath (40° C.) to dryness to yield a syrup5,6-dichloro-1-(2,3,5-tri-O-acetyl-β-D-ribofuranosyl)benzimidazole-2-thione!. The syrup was dissolved in methanolic ammonia(methanol saturated with ammonia at 0° C., 100 ml) and allowed to standat room temperature for 24 hours. After the methanolic ammonia had beenremoved at room temperature, a syrup remained which was triturated withcold water (30 ml). The solid material (2.548 g) which separated wascollected by filtration and was applied to a flash column chromatographyon silica gel with CH₂ Cl₂ -MeOH (15:1) as eluant. Fractions werecollected (25 ml each), and fractions containing the desired compoundwere combined and evaporated to obtain 53 which was crystallized fromaq. EtOH. Overall yield 2.0 g, 57.0%, mp 241° C.

2-Benzylthio-5,6-dichloro-1-(β-D-ribofuranosylbenzimidazole (54)

Compound 53 (0.5 g) was dissolved in cold water (10 ml) containingconcentrated ammonium hydroxide (2.5 ml). Benzyl chloride (0.5 ml) wasadded to this solution with stirring and the stirring continued at roomtemperature for 5 hours. The white solid (0.41 g) which had separatedfrom solution was collected by filtration, washed with cold water. Thesolid was extracted with ethyl acetate (100 ml) and the ethyl acetatephase was dried over anhydrous Na₂ SO₄ and then evaporated by aspiratorto give a syrup. The syrup was subjected to a flash column of silica gelwith CH₂ Cl₂ -MeOH (15:1) as eluant. Fractions were collected (25 mleach) and fractions containing the desired compound were combined andevaporated to afford 54 which was crystallized from CH₂ C1₂ -MeOH(15:1), mp. 149° C., Yield 0.223 g (35.5%).

2-Chloro-5,6-dibromo-1-(2,3,5-tri-O-acetyl-β-D-ribofuranosyl)benzimidazole(56) and2-Chloro-5,6-dibromo-1-(2,3,5-tri-O-acetyl-α-D-ribofuranosyl)benzimidazole

A suspension of 931 mg (3 mmole) of 2-chloro-5,6-dibromobenzimidazole(17) in 15 ml of dry CH₃ CN was treated with 0.8 mL (3 mmole) ofN,O-bis(trimethylsilyl) trifluroacetamide (BSTFA) at 75° C. for 10 minto give a clear solution. To this solution, was added 955 mg (3 mmole)of 1,2,3,5-tetra-O-acetyl-β-D-ribofuranose and 0 64 mL (3.3 mmole) oftrimethylsilyl triflate (TMSTf). Stirring was continued at 75° C. for 1hr. The reaction mixture was cooled to room temperature and then dilutedwith 100 mL of EtOAc. The EtOAc solution was washed with a sat. NaHCO₃solution C100 mL×2), sat. NaCl solution (100 mL), dried (Na₂ SO₄), andevaporated. The residue was chromatographed on a silica column (3×20 cm,eluted with CHCl₃). Evaporation of fractions 4-10 and recrystallizationof the residue from MeOH gave 946 mg (56%) of 56 as white crystals. mp140°-142° C.; MS role 565.9082 (0.9%, M⁺ =565.9091); ¹ H NMR (DMSO-d₆ )d 8.20 (s, 1,7-H), 8.11 (s, 1,4-H), 6.25 (d, 1, 1'-H), 5.55 (t, 1,2'-H),5.43 (dd, 1,3'-H), 4.47 (m, 2,5'-H. 4'-H), 4.37 (m, 1, 5"-H), 2.15,2.14, 2.02 (3 X s, 3 X, 3.3 X Ac). Evaporation of fractions 13-15 gave392 mg (23%) of the a anomer as a white foam. ¹ H NMR (DMSO-d₆) d 8.07,8.06 (2 X s, 2, 4-H, 7-H), 6.70 (d, 1, 1'-H), 5.71 (t, 1, 2'-H), 5.50(dd, 1, 3'-H), 4.81 (m, 1, 4'-H), 4.36 (dd, 1,5'-H), 427 (dd, 1, 5"-H).

2-chloro-5,6-dibromo-1-(β-D-ribofuranosyl)benzimidazole (57)

Method A (57)

A solution of 569 mg (1 mmole) of (56) in 20 mL of NH₃ /MeOH was stirredin a sealed flask at room temperature for 5 hr. Volatile materials wereremoved by evaporation. The residue was recrystallized from MeOH to give337 mg (2 crops, 76%) of 57. Its ¹ H NMR spectrum was identical to thatof (57) prepared by a direct bromination; see Method B.

Method B (57)

To a suspension of 570 mg (2 mmole) of2-chloro-1-(β-D-ribofuranosyl)-benzimidazole in 50 mL of H₂ O, was addeddropwise 50 mL of Br₂ /H₂ O over a period of 30 min. The reactionmixture was stirred at room temperature for an additional 3 hr. (Thoughthe reaction mixture remained as a suspension, the shape of the crystalschanged and TLC indicated a clean reaction). The reaction mixture wasallowed to stand in an ice bath for 30 min. and then filtered, The solidwas washed with portions of H₂ O until the washings were neutral. Thissolid was air dried and recrystallized from MeOH to give 757 mg ofproduct (57) (80%) as white crystalline needles. MP 126°-129° C.; MS(FAB) m/e 440.8862 (3%, MH⁺ =440.8852); ¹ H NMR (DMSO-d₆) d 8.68 (s, 1,7-H) 8.09 (s, 1, 4-H), 5.88 (d, 1, 1'-H), 5.51 (d, 1, 2'-OH), 5.40 (t,1, 5'-OH ), 5.30 (d,I, 3'-OH), 4.42 (m, 1, 2'-H), 4.14 (m, 1, 3'-H),4.01 (m, 1, 4'-H), 3.70 m, 2, 5'-H).

2-Chloro-5,6-difluoro-1-(2,3,5-tri-O-acetyl-β-D-ribofuranosyl)benzimidazole(64) and2-Chloro-5,6-difluoro-1-(2,3,5-tri-O-acetyl-α-D-ribofuranosyl)benzimidazole

To a suspension of 0.943 g (5 mmole) of 12 in 25 mL of ClCH₂ CH₂ Cl, wasadded 1.25 mL (5 mmole) of BSA at 75° C. The reaction mixture wasstirred at 75° C. for 30 min. To this solution, was added 1.75 g (5.5mmole) of 1,2,3,5-tetra-O-acetyl-β-D-ribofuranose and 1.07 mL (5.5mmole) of TMSOTf. Stirring was continued at 75° C. for 30 min. Thereaction mixture was cooled to room temperature, diluted with 100 mL ofCHCl₃, and extracted with sat. NaHCO₃ solution (100 mL×2) and sat. NaClsolution (100 mL). The CHCl₃ phase was dried (Na₂ SO₄) and evaporated.The residue was chromatographed on a silica column (3×25 cm, elutedsuccessively with 0.5%, 1% MeOH/CHCl₃). Evaporation of the appropriatefractions and recrystailization of the residue from MeOH gave 1.257 g (3crops, 56%) of 64 as Crystalline needles. MP 90°-91° C.; MS m/e 446.0694(5%, M⁺ =446.0692); ¹ H NMR (DMSO-d₆) d 7.93 (dd, 1,7-H, ³ J_(F-H) =10.5Hz, ⁴ J_(F-H) =7.5 Hz), 7.82 (dd, 1, 4-H, ³ J_(F-H) =10.5 Hz, ⁴ J_(F-H)=7.5 Hz), 6.23 (d, 1, 1'-H, J_(1'-2') =7.0 Hz), 5.55 (t, 1,2'-H,J_(2'-3') =7.0 Hz), 5.44 (m, 1, 3'-H, J₃₋₄ =4.5 Hz), 4.45 (m, 3, 4'-Hand 5'-H), 2.13, 2.10, 2.02 (3 ×s, 9, 3×Ac); ¹³ C NMR (DMSO-d₆) d169.95, 169.50, 169.18 (3 x OCOCH₃), 148.76, 148.62, 146.10, 145.93 (C5and C6, ¹ J_(F-C) =242 Hz), 140.16 (C2), 137.01 (C3a, ³ J_(F-C) =12 Hz),128.54 (C7a, ³ J_(F-C) =12 Hz), 107.31, 107.09 (C4, J_(F-C) =20 Hz),100.93, 100.66 (C7, J_(F-C) =24 Hz), 86.89 (Cl'), 79.41 (C4'), 70.40(C2'), 68.62 (C3'), 62.67 (C5'), 20.43, 20.28, 20.00 (3 x OCOCH₃).

Further elution and evaporation of the appropriate fractions gave 0.456g (20%) of the a anomer as a syrup. MS m/e 446.0680 (12%, M⁺ =446.0692);¹ H NMR (DMSO-d₆): d 7.75 (m, 2, 7-H and 4-H), 6.69 (d, 1, 1'-H,J_(1'-2') =4.0 Hz), 5.69 (t, 1, 2'-H, J_(2'-3') =5.0 Hz), 5.49 (dd, 1,3'-H, J_(3'-4') =7.0 Hz), 4.90 (m, 1, 4'-H), 4.37 (dd, 1, 5'-H,J_(5'-4') =3.5 Hz, J_(5'-5') =12.0 Hz), 4.26 (dd, 1, 5"-H, J_(5"-4')=5.5 Hz), 2.09, 2.03, 1.54 (3 x s, 9, 3 x Ac); ¹³ C NMR (DMSO-d₆): d170.05, 169.27, 168.34 (3 x OCOCH₃), 148.54, 148.37, 148.19, 145.88,145.71 145.54 (C5 and C6, ¹ J_(F-C) =241 Hz, ² J_(F-C) =16 Hz), 139.68(C2), 136.71 136.58 (C3a, ³ J_(F-C) =11 Hz), 129.45, 129.32 (C7a, ³J_(F-C) =12 Hz), 106.66, 106.44 (C4, ² J_(F-C) =20 Hz), 101.79, 101.52(C7, ² J_(F-C) =24 Hz), 86.43 (C1'), 78.16 (C4'), 70.97 (C2'), 70.39(C3'), 62.69 (C5'), 20.49, 20.09, 19.48 (3 x OCOCH₃).

2-Chloro-5,6-difluoro-1-(β-D-ribofuranosyl)benzimidazole (65)

Compound 64 (0.894 g, 2 mmole) was treated with 20 mL of NH₃ /MeOH in apressure bottle at room temperature for 3 hr. The reaction mixture wasevaporated and coevaporated with MeOH to give a solid. This wasrecrystallized from MeOH to give 0.573 g (3 crops, 89%) of 65 as acrystalline compound. MP˜215° C. (dec.); MS m/e 320.0385 (20%, M⁺=320.0375); ¹ H NMR (DMSO-d₆): d 8.34 (dd, 1,7-H, ³ J_(F-H) =1.5 Hz, ⁴J_(F-H) =7.5 Hz), 7.77 (dd, 1, 4-H, ³ J_(F-H) =11.0 Hz, J_(F-H) =7.5Hz), 5.88 (d, 1, 1'-H, J_(1'-2') =8.0 Hz), 5.50 (d, 1, 2'-OH, J_(2-2-OH)=6.5 Hz), 5.45 (t, 1, 5'-OH, J_(5'-5'OH) =4.5 Hz), 5.30 (d, 1, 3'-OH,J_(3'-3'OH) =4.5 Hz), 4.40 (m, 1, 2'-H, J_(2'-3') =5.5 Hz), 4.14 (m, 1,3'-H, J_(3'-4') =1.5 Hz), 4.01 (m, 1, 4'-H), 3.72 (m, 2, 5'-H, J_(5'-4')=2.5 Hz, J_(5'-5") =12.0 Hz); ¹³ C NMR (DMSO-d₆): d 148.53, 148.42,148.37, 148.26, 145.88, 145.76, 145.72, 145.60 (C5 and C6, ¹ J_(F-C)=241 Hz, ² J_(F-C) =16 Hz), 140.84 (C2), 137.16, 137.04 (C3a, ³ J_(F-C)=11 Hz), 128.55, 128.42 (C7a, ³ J_(F-C) =12 Hz), 106.77, 106.54 (C4, ²J_(F-C) =20 Hz), 102.12, 101.84 (C7, ² J_(F-C) =25 Hz), 89.10 (C1'),86.39 (C4'), 71.55 (C2'), 69.81 (C3'), 6.12 (C5').

2,5-Dichloro-6-fluoro-1-(2,3,5-tri-O-acetyl-β-D-ribofuranosyl)benzimidazole(2,5-β-anomer)&2,6-Dichloro-5-fluoro-1-(2,3,5-tri-O-acetyl-β-D-ribofuranosyl)benzimidazole(2,6-β-anomer)and 2,5-Dichloro-6-fluoro-1-(2,3,5-tri-O-acetyl-α-D-ribofuranosylbenzimidazole (2,5-α-anomer) &2,6-Dichloro-5-fluoro-1-(2,3,5-tri-O-acetyl-α-D-ribofuranosyl)benzimidazole(2,6-α-anomer)

To a suspension of 0.410 g (2.0 mmol) of 12 c in 10 mL of MeCN, wasadded 0.5 mL (2.0 mmol) of BSA. The reaction mixture was stirred at 75°C. for 10 min to give a clear solution. This solution was treated with0.70 g (2.2 mmol) of 1,2,3,5-tetra-O-acetyl-β-D-ribofuranose and 0.464mL (2.4 mmol) of TMSOTf at 75° C. for 30 min. The reaction mixture wascooled and diluted with EtOAc (50 mL). The EtOAc solution was washedwith sat. NaHCO₃ solution (50 mL×2), sat. NaCl solution (50 mL), dried(Na₂ SO₄), and evaporated. The residue was chromatographed on a silicacolumn (2.4×20 cm, eluted with CHCl₃ and 0.5% MeOH/CHCl₃). Evaporationof fractions 21-32 (20 mL per fraction) and recrystallization from MeOHgave 0.438 g (47%) of the β-anomers (2,5-β-anomer and 2,6-β-anomer) aswhite crystals. MP: 117°-119° C. MS: (El) m/e 462.0414 (7%, M⁺=462.0397). ¹ H NMR (DMSO-d₆): d 8.03 d, 0.4, 7-H (2,6-β-anomer), ⁴J_(F-H) =6.5 Hz!, 7.96 d, 0.6, 4-H (2,5-β-anomer), ⁴ J_(F-H) =7.0 Hz!,7.92 d, 0.6, 7-H (2,5-β-anomer), ³ J_(F-H) =9.5 Hz!, 7.80 d, 0.4, 4-H(2,6-β-anomer), ³ J_(F-H) =9.5 Hz!, 6.25 d, 0.4, 1'-H (2,6-β-anomer),J_(1'-2') =7.0 Hz!, 6.24 d, 0.6, 1'-H (2,5-β-anomer), J_(1'-2') =7.0Hz!, 5.55 t, 0.4, 2'-H (2,6-β-anomer), J_(2'-3') =7.0 Hz!, 5.54 t, 0.6,2'-H (2,5-β-anomer), J_(2'-3') =7.0 Hz!, 5.45 m, 1, 3'-H(2,5-β-anomer+2,6-β-anomer)!, 4.45 m, 3, 4'-H and 5'-H2,5-β-anomer+2,6-β-anomer)!, 2.14, 2.11, 2.02 3 x s, 9, 3 x Ac(2,5-β-anomer+2,6-`-anomer)!. Anal. Calcd. for C₁₈ H₁₇ Cl₂ FN₂ O₇ : C46.67, H 3.70, N 6.05. Found: C 47.76, H 3.66, N 5.97.

Evaporation of fractions 35-36 (20 mL per fraction) gave 0.153 g (17%)of the α-anomers (2,5-α-anomer and 2,6-α-anomer) as a foam. MS: (El) m/e462.0399 (9%, M⁺ =462.0397). ¹ H NMR (DMSO-d₆): d 7.88, 7.37 2 x twooverlapping d, 2, 4-H and 7-H (2,5-α-anomer+2,6-α-anomer), ³ J_(F-H)=9.5 Hz, ⁴ J_(F-H) =7.0 Hz!, 6.70 2 x d, 1, 1'-H(2,5α-anomer+2,6-α-anomer), J_(1'-2') =4.0 Hz!, 5.70 m, 1, 2'-H(2,5-α-anomer+2,6-α-anomer)!, 5.50 m, 1, 3'-H(2,5-α-anomer+2,6-α-anomer)!, 4.88 m, 1, 4'-H(2,5-α-anomer+2,6-α-anomer)!, 4.37 m, 1,5'-H(2,5-α-anomer+2,6-α-anomer)!, 4.27 m, 1, 5"-H(2,5-α-anomer+2,6-α-anomer)!, 2.090, 2.086, 2.043, 2.028, 1.548, 1.544 6x s, 9, 3 x Ac (2,5-α-anomer+2,6-α-anomer)!.

2,5-Dichloro-6-fluoro-1-β-D-ribofuranosylbenzimidazole 65a (2,5) &2,6-Dichloro-5-fluoro-1-β-D-ribofuranosylbenzimidazole 65a (2,6)

A solution of 0.323 g (0.50 mmol) of the β-anomer (2,5+2,6) in 10 mL ofNH₃ /MeOH was stirred in a pressure bottle at room temperature for 5 h.Volatile materials were removed by evaporation and coevaporation withMeOH (3 x, bath temperature<40° C.). The resulting solid wasrecrystallized from MeOH to give 0.141 g (2 crops, 84%) of whitecrystals (65a (2,5+2,6)). MP: 170°-172° C. MS: (El) m/e 336.0082 (14%,M⁺ =336.0080). ¹ H NMR (DMSO-d₆): d8.49 d, 0.4, 7-H (65a-2,6), ⁴ J_(F-H)=7.0 Hz!, 8.31 d, 0.6, 7-H (65a-2,5), ³ J_(F-H) =10.0 Hz!, 7.91 d, 0.6,4-H (65a-2,5), ⁴ J_(F-H) =7.0 Hz!, 7.74 d, 0.4, 4-H (65a-2,6), ³ J_(F-H)=9.5 Hz!, 5.89 d, 1, 1'-H (65a (2,5+2,6)), J_(1'-2') =7.0 Hz!, 5.50 d,1,2'-OH (65a (2,5+2,6)), J_(2'-2'OH) =6.5 Hz!, 5.43 2 x overlapping t,1, 5'-OH (65a (2,5+2,6)), J_(5'-5'OH) =4.5 Hz!, 5.29 d, 1, 3'-OH (65a(2,5+2,6)), J_(3'-3'OH) =4.5 Hz!, 4.41 m, 1, 2'-H (65a (2,5+2,6))!, 4.14m, 1, 3'-H (65a (2,5+2,6))!, 4.02 m, 1, 4'-H (65a (2,5+2,6))!, 3.70 m,2, 5'-H (65a (2,5+2,6))!. Anal. Calcd. for C₁₂ H₁₁ Cl₂ FN₂ O₄ : C,42,75; H, 3.29; N, 8.31. Found: C, 42.81; H, 3.43; N, 8.16.

2,6-Dichloro-1-(2,3,5-tri-O-acetyl-β-D-ribofuranosylbenzimidazole (66)

Method A (66)

6.24 grams (33.4 mmoles) of 35 was suspended in 150 ml of1,2-dichloroethane while stirring. To this material at 80° C., 8.5 ml(33.4 mmoles) of BSA was added via a syringe to give a clear solution.To this solution were added 10.62 grams (33.4 mmoles) of1,2,3,5-tetra-O-acetyl-β-D-ribofuranose and 7.0 ml (36.7 moles) ofTMSOTf. The reaction mixture was allowed to stir at 80° C. for one hr.EtOAc (100 ml) was added to the reaction mixture. The EtOAc solution wasextracted with saturated NaHCO₃ solution (300 ml×2), saturated NaClsolution (50 ml), dried (Na₂ SO₄), and evaporated in vacuo. Theresulting yellowish-brown syrup was dissolved in a minimum volume ofchloroform and transferred to a silica column (30×3 cm). The column waseluted with CRCl₃ while ultimately increasing solvent polarity to 1%MeOH/CHCl₃. The first spot eluted from the column was collected andevaporated in vacuo to reveal an oily yellow residue. The syrup wascoevaporated with EtOH (25 ml×2) to reveal a white precipitate.Recrystallization from MeOH afforded 4.47 grams (30.0%) of a mixture ofthe 2,5-dichloro and 2,6-dichloro isomers as colorless crystals.Fractional recrystallization yielded 2.815 grams (19%) of colorlesscrystalline 66. MP 154°-155° C. MS (El) m/e 444.0492 (6%, M⁺ =444.0491).¹ H NMR (DMSO-d₆): d 7.89 (d, 1, 7-H, J₇₋₅ =2.0 Hz), 7.68 (d, 1, 4-H,J₄₋₅ =8.5 Hz), 7.37 (dd, 1, 5-H), 6.25 (d, 1, 1'-H, J_(1'-2') =7.0 Hz),5.57 (t, 1, 2'H, J_(2'-3') =7.0 Hz), 5.44 (dd, 1, 3'-H, J_(3'-4') =4.5Hz), 4.47, 4.39 (2 x m, 3, 4'-H and 5'-H), 2.15, 2.14, 2.02 (3 x s, 9, 3x Ac). ¹³ C NMR (DMSO-d₆): d 169.94, 169.48, 169.15 (3 x OCOCH₃), 140.06(C3a and C2), 133.48 (C7a), 128.34 (C6), 123.78 (C5), 120.52 (C4),111.71 (C7), 86.65 (C1'), 79.40 (C4'), 70.35 (C2'), 68.64 (C3'), 62.58(C5'), 20.55, 20.26, 19.97 (3 x OCOCH₃).

Method B (66)

To a stirred mixture of 0.419 g (3.116 mmole) of CuCl₂ and 0.371 mL(2.807 mmole) of 90% t-BuONO in 5 mL of CH₃ CN, was added dropwisesolution of 0.633 g (1.487 mmole) of2-amino-6-chloro-1-(β-D-ribofuranosyl)benzimidazole in 3 mL of CH₃ CN.After the addition, stirring was continued at room temperature for 2 hr.The reaction mixture was diluted with 60 mL of EtOAc. The EtOAc solutionwas washed with H₂ O (50 mL), sat. NaHCO₃ solution (50 mL x 2), sat.NaCl solution (50 mL), dried (Na₂ SO₄), and evaporated. The residue waschromatographed on a silica column (1.9×35 cm, eluted with CHCl₃).Evaporation of the appropriate fractions and recrystallization from MeOHgave 0.380 g (57%) of 66 as a while cystalline compound. MP 155°-157° C.MS (El) m/e 444.0487 (7%,M⁺ =444.0491). ¹ H NMR (DMSO-d₆): d 7.89 (d, 1,7-H, J₇₋₅ =2.0 Hz), 7.68 (d, 1, 4-H, J₄₋₅ =8.5 Hz), 7.37 (dd, 1, 5-H),6.25 (d, 1, 1'-H, J_(1'-2') =7.0 Hz), 5.57 (t, 1, 2'-H, J_(2'-3') =7.0Hz), 5.45 (dd, 1, 3'-H, J_(3'-4') =4.5 Hz), 4.47, 4.39 (2 x m, 3, 4'-Hand 5'-H), 2.15, 2.14, 2.02 (3 x s, 9, 3 x Ac). ¹³ H NMR (DMSO-d₆) d169.91, 169,46, 169.12 (3 x OCOCH₃), 140.06, 140.02 (C3a and C2), 133.49(C7a), 128.34 (C6), 123.78 (C5), 120.51 (C4), 111.69 (C7), 86.65 (C1'),79.39 (C4'), 70.35 (C2'), 68.64 (C3'), 62.56 (C5'), 20.54, 20.25, 19.96(3 x OCOCH₃).

2,6-Dichloro-1-(β-ribofuranosyl)benzimidazole (67)

2,6-Dichloro-1-(2,3,5-tri-O-acetyl-β-D-ribofuranosyl)benzimidazole (66)(1.8 grams) was dissolved in methanolic ammonia (35 ml). The reactionmaterial was stirred in a pressure bottle for 5 hours at roomtemperature. Volatile materials were evaporated in vacuo to provide awhite powdery precipitate. This residue was recrystallized from MeOH toyield 561 mg (2 crops, 85%) of 67 as colorless needles. MP 162°-163° C.¹ H NMR (DMSO-d₆): d 8.30 (d, 1, 7-H, J₇₋₅ =2.0 Hz), 7.64 (d, 1, 4-H,J₄₋₅ =8.5 Hz), 7.31 (dd, 1, 5-H), 5.89 (d, 1, 1'-H, J_(1'-2') =8.0 Hz),5.49 (d, 1, 2'-OH, J_(2'-2') =6 5 Hz), 5.35 (t, 1, 5'-OH, J_(5'-5'OH)=5.0 Hz), 5.28 (d, 1,3'-OH, J_(3'-3'OH) =4.5 Hz), 4.46 (m, 1, 2'-H,J_(2'-3') =5.-5 Hz), 4.14 (m, 1,3'-H, J_(3'-4') =2.0 Hz), 4.00 (m,1,4'-H, J_(4'-5') =J_(4'-5') =3.0 Hz), 3.70 (m, 2, 5'H and 5"-H,J_(5'-5") =12.0 Hz). ¹³ C NMR (DMSO-d₆) d 140.72 C2), 140.17 (C3a),133.56 (C7a), 127.86 (C6), 123.23 (C5), 128.34 (C6), 120.03 (C4), 113.15(C7), 89.00 (C1'), 86.26 (C4'), 71.37 (C2'), 69.72 (C3'), 61.12 (C5').

2,5-Dichloro-1-(2,3,5-tri-O-acetyl-β-D-ribofuranosyl)benzimidazole (72)

To a stirred mixture of 0.292 g (2.172 mmole) of CuCl₂ and 0.259 mL(1.960 mmole) of 90% t-BuONO in 4 mL of CH₃ CN, was added dropwise asolution of 0.463 g (1.087 mmole) of2-amino-5-chloro-1-(β-D-ribofuranosyl)benzimidazole in 2 mL of CH₃ CN.After the addition, stirring was continued at room temperature for 2 hr.The reaction mixture was diluted with 50 mL of EtOAc. The EtOAc solutionwas washed with H₂ O (50 mL), sat. NaHCO₃ solution (50 mL x 2), sat.NaCl solution (50 mL), dried (Na₂ SO₄), and evaporated. The residue waschromatographed on a silica column (2×15 cm, eluted with CHCl₃).Evaporation of the appropriate fractions and recrystallization from MeOHgave 0.265 g (55%) of 72 as a white crystalline compound. MP 98°-100° C.MS (El) m/e 444.0487 (8%, M⁺ =444.0491). ¹ H NMR (DMSO-d₆): d 7.81 (d,1, 7-H, J₇₋₆ =9.0 Hz), 7.77 (d, 1, 4-H, J₄₋₆ =2.0 Hz), 7.42 (dd, 1,6-H), 6.25 (d, 1, 1'-H, J_(1'-2') =7.0 Hz), 5.57 (t, 1, 2'-H, J_(2'-3')=7.0 Hz), 5.42 (dd, 1, 3'-H, J_(3'-4') =5.0 Hz), 4.43 (m, 3, 4'-H and5'-H), 2.13, 2.11, 2.02 (3 x s, 9, 3 x Ac). ¹³ H NMR (DMSO-d₆): d170.01, 169.52, 169.20 (3 x OCOCH₃), 142.17 (C3a), 140.66 (C2), 131.73(C7a), 127.94 (C5), 123.91 (C6), 118.80 (C4), 113.19 (C7), 86.88 (C1'),79.26 (C4'),70.46 (C2'), 68.73 (C3'), 62.65 (C5'), 20.52, 20.29, 20.01(3 x OCOCH₃).

2,5-Dichloro-1-(β-D-ribofuranosyl)benzimidazole (73)

A solution of 0.226 g (0.508 mmole) of 72 in 10 mL of NH₃ /MeOH wasstirred in a pressure bottle at room temperature for 5 hr. The reactionmixture was evaporated and coevaporated with MeOH (3 x) to give asolid). This solid was recrystallized from MeOH to give 0.136 g (76%,based on C₁₂ H₁₂ Cl₂ N₂ O₄ MeOH) of 73 as white crystals. MP 102°-150°C. (melted over a large range of temperature). MS (Cl) m/e 319.0242(37%, MH⁺ =319.0252). ¹ H NMR (DMSO-d₆): d 8.06 (d, 1, 7-H, J₇₋₆ =9.0Hz), 7.73 (d, 1, 4-H, J₄₋₆ =2.0 Hz), 7.30 (dd, 1, 6H), 5.89 (d, 1, 1'-H,J₁₋₂ =8.0 Hz), 5.51 (d, 1, 2'-OH, J_(2'-2'OH) =6-5 Hz), 5.29 (d, 1,3'-OHJ_(3') -_(3'OH) =4.5 Hz), 5.26 (t, 1, 5'-OH, J_(5'-5'OH) =5-0 Hz), 4.45(m, 1, 2'-H, J_(2'-3') =5.5 Hz), 4.12 (m, 1, 3'-H, J_(3'-4') =2.0 Hz),3.99 (m, 1, 4'-H, J_(4'-4') =J_(4'-5") =3.5 Hz), 3.69 (m, 2, 5'-H and5"-H, J_(5'-5") =12.0 Hz). ¹³ C NMR (DMSO-d₆) d 142.36 (C3a), 141.40(C2), 131.88 (C7a), 127.49 (C5), 123.32 (C6), 118.40 (C4), 114.51 (C7),89.07 (C1'), 86,23 (C4'), 71.49 (C2'), 69.75 (C3'), 61.23 (C5').

2-Chloro-5-nitro-1-(2,3,5-tri-O-acetyl-β-D-ribofuranosyl)benzimidazole(74) and2-Chloro-6-nitro-1-(2,3,5-tri-O-acetyl-β-D-ribofuranosyl)benzimidazole(75)

To a suspension of 3.952 g (20 mmole) of 19 in 100 mL of1,2-dichloroethane, was added 5 mL (20 mmole) of BSA. The reactionmixture was stirred at 75° C. for 15 min to give a clear solution. Thissolution was cooled to ˜20° C. and treated with 7.0 g (22 mmole) of1,2,3,5-tetra-O-acetyl-b-D-ribofuranose and 4.638 mL (24 mmole) ofTMSOTf at room temperature for 2 h. The reaction mixture was dilutedwith 200 mL of CHCl₃. The CHCl₃ solution was washed with sat. NaHCO₃solution (200 mL x 2), sat. NaCl solution (200 mL) dried (Na₂ SO₄), andevaporated. The residue was chromatographed on a silica column (5×35 cm,eluted with CHCl₃ and 0.5% MeOH/CHCl₃). Evaporation of the appropriatefractions gave 6.50 g (71%. one spot on TLC) of 74 and 75 as a whitefoam. Fractional recrystallization of this foam (5 times from MeOH) give1.59 g (17%) of the pure 6-nitro isomer 75. MP 127°-129° C. MS (El) m/e455.0750 (2%, M⁺ =455.0732). ¹ H NMR (DMSO-d₆): d 8.68 (d, 1, 7-H, J₇₋₅=2.0 Hz), 8.21 (dd, 1, 5-H, J₅₋₄ =9.0 Hz), 7.88 (d, 1, 4-H), 6.41 (d, 1,1'-H, J_(1'-2') =7.0 Hz), 5.58 (t, 1, 2'-H, J_(2'-3') =7.0 Hz), 5.45(dd, 1, 3'-H J_(3'-4') =4.0 Hz), 4.50, 4.40 (2 x m, 3, 4'-H and 5'-H),2.15, 2.12, 2.03 (3 x, s, 9, 3 x Ac). ¹³ C NMR (DMSO-d₆): 170.12,169.49, 169.25 (3 x OCOCH₃), 145.60 (C3a), 143.96 (C2), 143.65 (C6),132.46 (C7a), 119.65 (C4), 118.92 (C5), 108.47 (C7), 86.96 (C1'), 79.67(C4'), 71.12 (C2') 68.75 (C3'), 62.56 (C5'), 20.39, 20.31, 20.03 (3 xOCOCH₃).

5-Amino-2-chloro-1-(2,3,5-tri-O-acetyl-β-D-ribofuranosyl)-benzimidazole(76) and6-Amino-2-chloro-1-(2,3,5-tri-O-acetyl-β-D-ribofuranosyl)benzimidazole(77)

A sample of the foam containing 74 and 75 (3.96 g, 8.688 mmole, one spoton TLC) was dissolved in 90 mL of EtOH and was hydrogenated at roomtemperapture, 50 psi for 1 day using ˜0.30 g of Raney Nickel as catalyst(the reaction was monitored by TLC). The reaction mixture was thenfibred and the filtrate was evaporated. The residue was chromatographedon a silica column (5×45 cm, eluted with 0.5% MeOH/CHCl₃). Evaporationof the appropriate fractions gave 1.54 g (42%) of 77 as a white foam. MS(El) m/e 425.0987 (14%, M⁺ =425.0990). ¹ H NMR (DMSO-d₆): d 7.29 (d, 1,4-H, J₄₋₅ =8.5 Hz). 6.75 (d, 1, 7-H, J₇₋₅ =1.5 Hz), 6.61 (dd, 1, 5-H),6.09 (d, 1, 1'-H, J_(1'-2') =6.5 Hz), 5.57 (t, 1, 2'-H, J_(2'-3') =7-5Hz), 5.37 (dd, 1, 3'-H, J_(3'-4') =5.5 Hz), 5.19 (s, 2, 6-NH₂), 4.43(dd, 1,5'-H, J_(5'-4') =2.5 Hz, J_(5'-5") =11.0 Hz), 4.40 (m, 1, 4'-H),4.36 (dd, 1, 5"-H, J_(5"-5') =6.0 Hz), 2.13, 2.09, 2.04 (3 x s, 9, 3 xAc). ¹³ C NMR (DMSO-d₆); d 170.10, 169.45, 169.19 (3 x OCOCH₃), 146.03(C6), 134.83 (C2), 134.06 (C7a), 132.92 (C3a), 119.35 (C4), 112.17 (C5),94.72 (C7), 86.67 (C1'), 78.46 (C4'), 69.75 (C2'), 68.58 (C3'), 62.51(C5'), 20.50, 20.21, 19.99 (3 x OCOCH₃).

Further elution and evaporation of the appropriate fractions gave 1.86 g(50%) of 76 as a white foam. MS (El) m/e 425.0976 (9%, M⁺ =425.0990). ¹H NMR (DMSO-d₆): d 7.40 (d, 1, 7-H, J₇₋₆ =8.5 Hz), 6.76 (d, 1,4-H, J₄₋₆=2.0 Hz), 6.66 (dd, 1,6-H, 6.10 (d, 1, 1'-H, J_(1'-2') =6.5 Hz), 5.56(t, 1, 2'-H, J_(2'-3') =7.0 Hz), 5.39 (dd, 1, 3'-H, J_(4'-4') =5.0 Hz),5.10 (br. s, 2, 5-NH₂), 4.40 (m, 3, 4'-H and 5'-H), 213, 2.10, 2.02 (3 xs, 9, 3 x Ac). ¹³ C NMR (DMSO-d₆): d 169.91, 169.52, 169.11 (3 xOCOCH₃), 145.31 (C5), 142.74 (C3a), 137.80 (C2), 124.66 (C7a), 112.50(C6), 111.62 (C7), 102.23 (C4), 86.57 (C1'), 78.89 (C4'), 70.05 (C2'),68.78 (C3'), 62.69 (C5'), 20.45, 20.26, 19.98 (3 x OCOCH₃).

5-Amino-2-chloro-1-(β-D-ribofuranosyl)benzimidazole (78)

A solution of 0.363 g (0.852 mmole) of 76 in 10 mL of NH₃ /MeOH wasstirred in a pressure bottle at room temperature for 5 hr. The reactionmixture was evaporated and coevaporated with MeOH (3 x, bath temperature<40° C.). The residue was recrystallized from MeOH/Et₂ O to give 0.203 g(2 crops, 79%) of 78 as beige crystals. MP ˜154° C. (dec.). MS (El) m/e299.0675 (10%, M⁺ =299.0673). ¹ H NMR (DMSO-d₆): d 7.56 (d, 1, 7-H, J₇₋₆=8.5 Hz), 6.73 (d, 1,4-H, J₄₋₆ =2.0 Hz), 6.57 (dd, 1, 6-H), 5.77 (d, 1,1'-H, J_(1'-2') =7 5 Hz), 5.39 (d, 1, 2'-OH, J_(2'-2'OH) =6.5 Hz), 5.18(d, 1, 3'-OH, J_(3'-3'OH) =4.5 Hz), 5.11 (t, 1, 5'-OH, J_(5'-5'OH) =5.0Hz), 4.90 (s, 2, 5-NH₂), 4.44 (m, 1, 2'-H, J_(2'-3') =6.0 Hz), 4.09 (m,1, 3'-H, J_(3'-4') =2.5 Hz), (3.91 (m, 1, 4'-H, J_(4'-5') =3.5 Hz), 3.65("t", 2, 5'-H). ¹³ C NMR (DMSO-d₆): d 144.82 (C5), 142.77 (C3a), 138.25(C2), 125.13 (C7a), 112.67 (C7), 112.22 (C6), 101.83 (C4), 88.81 (C1'),85.48 (C4'), 70.89 (C2'), 69.61 (C3'), 61.31 (C5').

6-Amino-2-chloro-1-(β-D-ribofuranosyl)benzimidazole (79)

A solution of 0.283 g (0.665 mmole) of 77 in 10 mL of NH₃ /MeOH wasstirred in a pressure bottle at room temperature for 5 hr. The reactionmixture was evaporated and coevaporated with MeOH (3 x, bath temperature<40° C.). The residue was recrystallized from MeOH to give 0.170 g (2crops, 85%) of 79 as beige crystals. MP ˜170° C. (dec.). MS (El) m/e299.0862 (16%, M⁺ =299.0673). ¹ H NMR (DMSO-d₆): d 7.25 (d, 1,4-H, J₄₋₅=8.5 Hz), 6.87 (d, 1, 7-H, J₇₋₅ =2.0 Hz), 6.57 (dd, 1, 5-H), 5.75 (d, 1,1'-H, J_(1'-2') =7 5 Hz), 5.43 (d, 1,2'-OH, J_(2'-2'OH) =6-5 Hz), 5.18(d, 1, 3'-OH, J_(3'-3'OH) =5.0 Hz), 5.04 (s, 2, 6-NH₂), 5.03 (t, 1,5'-OH, J_(5'-5'OH) =5.5 Hz), 4.48 (m, 1, 2'-H, J_(2'-3') =6.0 Hz), 4.07(m, 1,3'-H, J_(3'-4') =3.0 Hz), 3.90 (m, 1, 4'-H, J_(4'-5') =4.0 Hz,J_(4'-5') =5.0 Hz), 3.66 (m, 2, 5'-H and 5"-H, J_(5'-5") =12.0 Hz). ¹³ CNMR (DMSO-d₆): d 145.34 (C6), 135.63 (C2), 134.34 (C7a), 133.18 (C3a),118.83 (C4), 111.71 (C5), 95.79 (C7), 88.79 (C1'), 85.24 (C4'), 70.17(C2'), 69.70 (C3'), 61.55 (C5').

4,6-Dichloro-2-trifluoromethyl-1-(2,3,5-tri-O-acetyl-1-β-D-ribofuranosyl)benzimidazole(81a)

A mixture of 4,6-dichloro-2-trifluoromethylbenzimidazole (28a, 0.99 g,3.4 mmole) and 1,2,3,5-tetra-O-acetyl-β-D-ribofuranose (1.71 g, 5.36mmole) was heated at 170° C. for 1.5 hr under reduced pressure. Theresulting mixture was dissolved in 20 mL of CHCl₃ and waschromatographed on a SiO₂ column, (2×20 cm). Elution of the column withhexane:EtOAc (80:20, v/v) and evaporation of appropriate fraction gave0.67 gm (34%) of 81a: m.p. 116°-120° C; ¹ H NMR (DMSO-d₆): d 8.00 (d,1H, J=1.6 Hz, 7H), 7.74 (d, 1H, J=1.5 Hz, 5H), 6.15 (d, 1H, J_(1'-2')=6.8 Hz, 1'H), 5.53 (t, 1H, J_(2'-3') =6.7 Hz, 2'H), 5.49-5.45 (m, 1H,3'H), 4.56-4.53 (m, 2H, 4'H and 5'H), 4.37-4.33 (m, 2H, 5"H), 21.5,2.12, 1.92 (3s, 9H, 3 x Ac).

4,6-Dichloro-2-trifluoromethyl-1-(β-D-ribofuranosyl)benzimidazole (81b)

A mixture of 81a (0.57 g, 1.1 mmole) and MeOH-NH₃ (20 mL) were stirredat room temperature for 10 hr. The excess of MeOH and NH₃, was removedunder reduced pressure. The product thus obtained was chromatographedover SiO₂ column (2×14 cm) (230-400 mesh). Elution of the column withCHCl₃ :MeOH (98:2, v/v) the product which was crystallized from themixture of MeOH-H₂ O to give 0.268 g (62.4%) of 81b: m.p. 162° C.; ¹ HNMR (DMSO-d₆): d 8.59 (d, 1H, J=1.7 Hz, 7H), 7.68 (d, 1H, J=1.7 Hz, 5H),5.82 (d, 1H, J_(1-2') =7.62 Hz, 1'H), 5.50-5.47 (m, 2H, exchanges withD₂ O, 2'OH and 5'OH), 5.29 (d, 1H, exchanges with D₂ O), 4.41 (q, 1H,J_(2'-3') =6.06 Hz, 2'H), 4.16 (bs, 1H, 3'H), 4.04 (bs, 1H, 4'H), 3.75(m, 2H, S'H and 5"H). Anal. Calcd, for (C₁₃ H₁₁ N₂ O₄ F₃ Cl₂): C, 40.33;H, 2.86; N, 7.23; Found: C, 40.02; H, 2.66; N, 6.74.

2,4-Dichloro-1-(2,3,5-tri-O-benzyl-β-D-ribofuranosyl)-6-trifluoromethytbenzimidazoleand2,4-Dichloro-1-(2,3,5-tri-O-benzyl-α-D-ribofuranosyl)-6-trifluoromethylbenzimidazole

For the preparation of 1-O-chloro-2,3,5-tri-O-benzyl-D-ribofuranose, HClgas was allowed to pass through a solution of1-O-p-nitrobenzoyl-2,3,5-tri-O-benzyl-β-D-ribofuranose (0.774 g, 1.36mmol) in 5 mL of dry CH₂ Cl₂ at 0° C. for 15 min. The resultingsuspension was quickly filtered and the filtrate was evaporated. Theresidue was coevaporated with MeCN and then dissolved in 5 mL of dryMeCN or immediate use in the subsequent glycosylation reaction.

To a mixture of 0.289 g (1.133 mmol) of 41 c in 5 mL of MeCN, was added0.283 mL (1.133 mmol) of BSA. The reaction mixture was stirred at 80° C.for 25 min. This solution was reacted with the above MeCN solution ofthe carbohydrate and 0.285 mL (1.473 mmol) of TMSOTf at 80° C. for 30min. The reaction mixture was cooled and diluted with EtOAc (60 mL). TheEtOAc solution was washed with sat. NaHCO₃ solution (50 mL×2); sat. NaClsolution (50 mL), dried (Na₂ SO₄), and evaporated. The residue waschromatographed on a silica column (1.9×35 cm, eluted with 10%, 15%, 20%EtOAc/hexane). Evaporation of fractions 20-26 (20 mL per fraction) gave0.31 g (42%) of the β-anomer as a syrup. MS: (Cl) m/e 657.1561 (1%, MH⁺=657.1535). ¹ H NMR (DMSO-d₆): d 8.05 (s, 1, 7-H), 7.69 (s, 1, 5-H),7.35, 6.95 (2 x m, 15, 3 x Ph) 6.08 (d, 1, 1'H, J_(1'-2') =8.0 Hz),4.73˜4.24 (m, 9, 2'-H, 3'-H, 4'-H, and 3 x PhCH₂), 3.75 (dd, 1, 5'-H,J_(4'-5') =2.0 Hz, J_(5'-5") =11.0 Hz), 3.58 (dd, 1, 5"-H, J_(4'-5")=3.0 Hz).

Evaporation of fractions 34-41 (20 mL per fraction) gave 0.262 g (35%)of the a-anomer as a syrup. MS: (Cl) m/e 657.1517 (1%, MH⁺ =657.1535). ¹H NMR (DMSO-d₆): d 7.94 (s, 1, 7-H), 7.66 (s, 1, 5-H), 7.30, 7.08, 6.72(3 x m, 15, 3 x Ph), 6.51 (d, 1, 1'-H, J_(1'-2') =4.5 Hz), 4.69˜4.07 (m,9, 2'-H, 3'-H, 4'-H, and 3 x PhCH₂), 3.73 (dd, 1, 5'-H, J_(4'-5') =2.5Hz, J_(5'-5") =11.0 Hz), 3.62 (dd, 1, 5"-H, J_(4'-5") =4.5 Hz).

2,4-Dichloro-1-(β-D-ribofuranosyl)-6-trifluoromethylbenzimidazole (81 c)

To a solution of 0.29 g (0.441 mmol) of the β-anomer (see previousprep.) in 6 mL of CH₂ Cl₂, was added dropwise 4.41 mL of 1M BCl₃ at -78°C. The reaction mixture was stirred at -78° C. for 2 hr and then at -40°C. for 2 hr MeOH (3 mL) was added and stirring was continued at -40° C.for 10 min. The reaction mixture was diluted with EtOAc (50 mL). TheEtOAc solution was washed with H₂ O (50 mL), sat. NaHCO₃ solution (50mL), sat. NaCl solution (50 mL), dried (Na₂ SO₄), and evaporated. Theresidue was suspended in a small amount of CHCl₃ and then filtered. Thesolid product was washed with portions of CHCl₃ and dried in vacuo togive 0.146 g (86%) of 81 c as a white solid. MS: (El) m/e 386.0038 (14%,M⁺ =386.0048). ¹ H NMR (DMSO-d₆): d 8.75 (s, 1,7-H), 7.76 (s, 1, 5-H),5.97 (d, 1, 1'-H, J_(1'-2') =8.0 Hz), 5.55 (d, 1, 2'-OH, J_(2'-2'OH)=6.0 Hz), 5.43 (t, 1, 5'-OH, J_(5'-5'OH) =4.5 Hz), 5.35 (d, 1, 3'-OH,J_(3'-3'OH) =4.5 Hz), 4.43 (m, 1, 2'-H, J_(2'-3') =5.5 Hz), 4.15 (m, 1,3'-H, J_(3'-4') =1.5 Hz), 4.06 (m, 1, 4'-H, J_(4'-5') =J_(4'-5") =2.5Hz), 3.69 (m, 2, 5'-H and 5"-H).

2-Chloro-4,5-dibromo-1-(2,3,5-tri-O-acetyl-β-D-ribofuranosyl)benzimidazole(84)and2-Chloro-5-bromo-1-(2,3,5-tri-O-acetyl-β-D-ribofuranosyl)benzimidazole(86)

To a stirred mixture of 1.228 g (5.498 mmole) of CuBr₂ and 0.654 mL(4.949 mmole) of 90% t-BuONO in 10 mL of CH₃ CN, was added dropwise asolution of 1.170 g (2.748 mmole) of 78 in 3 mL of CH₃ CN. After theaddition, stirring was continued at room temperature for 2 hr. Thereaction mixture was diluted with 100 mL of EtOAc. The EtOAc solutionwas washed with H₂ O (100 mL), sat. NaHCO₃ solution (100 mL×2), sat.NaCl solution (100 mL), dried (Na₂ SO₄), and evaporated. The residue waschromatographed on a silica column (4.1×30 cm, eluted with CHCl₃).Evaporation of fractions 59-78 (20 mL per fraction) andrecrystallization from MeOH gave 0.600 g (38%) of 84 as a white solid.MP 202°-203° C. MS (El) m/e 565.9109 (4%,M⁺ =565.9091). ¹ H NMR(DMSO-d₆): d 7.74 (2 x d, 2, 6-H and 7H, J₆₋₇ =8.5 Hz), 6.26 (d, 1,1'-H, J_(1'-2') =6.5 Hz), 5.53 (t, 1, 2'-H, J_(2'-3') =7.0 Hz), 5.42(dd, 1, 3'-H, J_(3'-4') =4.5 Hz), 4.42 (m, 3, 4'-H and 5'-H), 2.14,2.11, 2.01 (3 x s, 9, 3 x Ac). ¹³ C NMR (DMSO-d₆): d 169.92, 169.42,169.14 (3 x OCOCH₃), 141.56 (C3a), 140.91 (C2), 132.12 (C7a), 128.03(C6), 118.58 (C5), 114.56 (C4), 112.60 (C7), 87.12 (C1'), 79.41 (C4'),70.60 (C2'), 68.66 (C3'), 62.55 (C5'), 20.47, 20.22, 19.94 (3 x OCOCH₃).

Evaporation of fractions 89-125 (20 mL per fraction) andrecrystallization from MeOH gave 0.405 g (30%) of 86 as a whitecrystals. MP 129°-130° C. MS (El) m/e 487.9973 (5%, M⁺ =487.9986). ¹ HNMR (DMSO-d₆) d 7.91 (d, 1, 4-H, J₄₋₆ =2.0 Hz), 7.76 (d, 1,7-H, J₇₋₆=8.5 Hz), 7.53 (dd, 1, 6-H), 6.25 (d, 1, 1'-H, J_(1'-2') =6-5 Hz), 5.56(t, 1, 2'-H, J_(2'-3') =7.0 Hz), 5.42 (dd, 1, 3'-H, J_(3'-4') =5.0 Hz),4.43 (m, 3, 4'-H and 5'-H), 2.14, 2.11, 2.02 (3 x s, 9, 3x Ac). ¹³ C NMR(DMSO-d₆): d 170.01, 169.52, 169.19 (3 x OCOCH₃), 142.61 (C3a), 140.50(C2), 132.05 (C7a), 126.51 (C6), 121.73 (C4), 115.71 (C5), 113.58 (C7),86.87 (C1'), 79.26 (C4'), 70.44 (C2'), 68.73 (C3'), 62.64 (C5'), 20.53,20.29, 20.01 (3 x OCOCH₃).

5-Bromo-2-chloro-1-(β-D-ribofuranosylbenzimidazole (87)

A solution of 0.304 g (0.621 mmole) of 86 in 10 mL of NH₃ /MeOH wasstirred in a pressure bottle at room temperature for 5 h. The reactionmixture was evaporated and coevaporated with MeOH (3 x, bath temperature<40° C.). The residue was recrystallized from MeOH to give 0.192 g (85%)of 87 as white crystals. MP 154°-155° C. MS (Cl) m/e 362.9735 (40%, MH⁺=362.9747). ¹ H NMR (DMSO-d₆): d 8.01 (d, 1, 7-H, J₇₋₆ =8.5 Hz), 7.87(d, 1, 4-H, J₄₋₆ =2.0 Hz), 7.41 (dd, 1, 6H), 5.89 (d, 1, 1'-H, J_(1'-2')=8.0 Hz), 5.51 (d, 1, 2'-OH, J_(2'-2'OH) =6-5 Hz), 5.28 (d, 1,3'-OH,J_(3'-3'OH) =4 5 Hz), 5.25 (t, 1, 5'-OH, J_(5'-5'OH) =5.0 Hz), 4.44 (m,1, 2'-H, J_(2'-3') =5.5 Hz), 4.13 (m, 1, 3'-H, J_(3'-4') =2.5 Hz), 3.98(m, 1, 4'-H, J_(4'-5') =J_(4'-5") =3.5 Hz), 3.68 (m, 2, 5'-H and 5"-H,J_(5'-5") =12.0 Hz). ¹³ C NMR (DMSO-d₆): d 142.81 (C3a), 141.23 (C2),132.20 (C7a), 125.92 (C6), 121.32 (C4), 115.27 (C5), 114.89 (C7), 89.08(C1'), 86.21 (C4'), 71.47 (C2'), 69.73 (C3'), 61.22 (C5').

2-Chloro-6,7-dibromo-1-(2,3,5-tri-O-acetyl-β-D-ribofuranosyl)-benzimidazole(88) and2-Chloro-6-bromo-1-(2,3,5-tri-O-acetyl-β-D-ribofuranosyl)benzimidazole(89)

To a stirred mixture of 0.564 g (2.525 mmole) of CuBr₂ and 0.300 mL(2.270 mmole) of 90% t-BuONO in 5 mL of CH₃ CN, was added dropwise asolution of 0.538 g (1.263 mmole) of 77 in 3 mL of CH₃ CN. After theaddition, stirring was continued at room temperature for 2 hr. Thereaction mixture was diluted with 60 mL of EtOAc. The EtOAc solution waswashed with H₂ O (50 mL), sat. NaHCO₃ solution (50 mL×2), sat. NaClsolution (50 mL), dried (Na₂ SO₄), and evaporated. The residue waschromatographed on a silica column (1.9×45 cm, eluted with CHCl₃).Evaporation of fractions 11-22 (20 mL per faction) and recrystallizationfrom MeOH twice gave 0.198 g of 89 as white crystals. Evaporation offractions 23-26 and recrystallization from MeOH gave 0.015 g of 88. Themother liquors were evaporated and the residue was rechromatographed ona silica column (1.9×45 cm, eluted with CHCl₃). Evaporation of fractions24-34 and recrystallization from MeOH twice gave an additional 0.072 gof 89 as white crystals. Fractions 37-56 were evaporated and the residuewas repurified by repeating the column chromatography. Evaporation ofthe appropriate fractions and recrystallization from MeOH gave anadditional 0.070 g of 88 as white crystals. The total yield of 88 was0.085 g (12%). MP 121°-122° C. MS (El) m/e 565.9092 (1%, M⁺ =565.9091).¹ H NMR (DMSO-d₆): d 7.74 (d, 1, 5-H, J₅₋₄ =8.5 Hz), 7.65 (d, 1, 4-H),7.25 ("br. s", 1, 1'-H), 5.80 (dd, 1, 2'-H, J_(2'-1') =5.0 Hz, J_(2'-3')=7.5 Hz), 5.44 (t, 1, 3'-H, J_(3'-4') =7.5 Hz), 4.46 (dd, 1, 5'-H,J_(5'-4') =3.0 Hz, J_(5'-5') =12.0 Hz), 4.39 (m, 1, 4'-H), 4.29 (dd, 1,5"-H, J_(5"-4") =6.0 Hz), 2.13, 2.08, 2.05 (3 x s, 9, 3x Ac). ¹³ C NMR(DMSO-d₆): d 169.97, 169.45, 169.33 (3 x OCOCH₃), 141.51 (C3a), 141.09(C2), 133.28 (C7a), 128.37 (C5), 121.92 (C6), 120.07 (C4), 105.20 (C7),86.58 (C1'), 78.06 (C4'), 71.34 (C2'), 68.24 (C3'), 62.08 (C5'), 20.43,20.08, 20.01 (3 x OCOCH₃). The total yield of 89 was 0.270 g (44%). MP169°-170° C. MS (El) m/e 487.9973 (9%, M⁺ =487.9986). ¹ H NMR (DMSO-d₆):d 8.01 (d, 1, 7-H, J₇₋₅ =1.5 Hz), 7.63 (d, 1, 4-H, J₄₋₅ =8.5 Hz), 7.49(m, 1, 5-H), 6.25 (d, 1, 1'-H, J_(1'-2') =7.0 Hz), 5.57 (t, 1, 2'-H,J_(2'-3') =7.0 Hz), 5.44 (dd, 1, 3'-H, J_(3'-3') =4.5 Hz), 4.47, 4.39 (2x m, 3, 4'-H and 5'-H), 2.16, 2.14, 2.02 (3 x s, 9, 3 x Ac). ¹³ C NMR(DMSO-d₆): d 169.99, 169.52, 169.19 (3 x OCOCH₃), 140.38 (C3a), 139.98(C2), 133.97 (C7a), 126.52 (C5), 120.91 (C4), 116.33 (C6), 114.50 (C7),86.61 (C1'), 79.43 (C4'), 70.39 (C2'), 68.66 (C3'), 62.61 (C5'), 20.71,20.31, 20.02 (3 x OCOCH₃).

6-Bromo-2-chloro-1-(β-D-ribofuranosyl)benzimidazole (90)

A solution of 0.176 g (0.359 mmole) of 89 in 10 mL of NH₃ /MeOH wasstirred in a pressure bottle at room temperature for 5 hr. The reactionmixture was evaporated and coevaporated with MeOH (3 x, bath temperature<40°C.). The resulting solid was recrystallized from MeOH to give 0.097g (2 crops, 74%) of 90 as white crystals. MP 152°-153° C. MS (El) m/e361.9685 (10%, M⁺ =361.9669). ¹ H NMR (DMSO-d₆): d 8.43 (d, 8.43 (d, 1,7-H, J₇₋₅ =2.0 Hz), 7.59 (d, 1, 4-H, J₄₋₅ =8.5 Hz), 7.43 (dd, 1, 5-H),5.88 (d, 1, 1'-H, J_(1'-2') =8.0 Hz), 5.49 (d, 1,2'-OH, J_(2'-2'OH) =6.5Hz), 5.34 (t, 1, 5'-OH, J_(5'-5'OH) =4.5 Hz), 5.27 (d, 1,3'-OH,J_(3'-3'OH) =4.5 Hz), 4.45 (m, 1,2'-H, J_(2'-3') =5.5 Hz), 4.14, (m,1,3'-H, J_(3'-4') =2.0 Hz), 4.00 (m, 1,4'-H, J_(4'-5'') =J_(4'-5") =3.5Hz), 3.70 (m, 2,5'-H and 5"-H, J_(5'-5") =12.0 Hz). ¹³ C NMR (DMSO-d₆):d 140.70 (C2), 140.49 (C3a), 134.02 (C7a), 125.95 (C5), 120.46 (C4),116.01 (C7), 115.87 (C6), 88.99 (C1'), 86.32 (C4'), 71.40 (C2'), 69.79(C3'), 61.15 (C5').

1-(2,3,5-Tri-O-benzyl-β-D-ribofuranosyl)-2,4,5,6-tetrachlorobenzimidazole(91a) and1-(2,3,5-Tri-O-benzyl-α-D-ribofuranosyl)-2,4,5,6-tetrachlorobenzimidazole

For the preparation of 1-O-chloro-2,3,5-tri-O-benzyl-D-ribofuranose, HClgas was allowed to pass through a solution of1-O-p-nitrobenzoyl-2,3,5-tri-O-benzyl-β-D-ribofuranose (1.479 g, 2.6mmol) in 10 mL of dry CH₂ Cl₂ at 0° C. for 15 min. The resultingsuspension was quickly filtered and the filtrate was evaporated. Theresidue was coevaporated with dry MeCN and then dissolved in 5 mL of dryMeCN for immediate use in the subsequent glycosylation reaction. To amixture of 13 (0.512 g, 2 mmol) in 10 mL of dry MeCN, was added 0.5 mL(2 mmol) of BSA. The reaction mixture was stirred at 75° C. for 20 min.This solution was treated with the above MeCN solution of the blockedcarbohydrate and 0.464 mL (2.4 mmol) of TMSOTf at 70° C. for 20 min. Thereaction mixture was cooled and diluted with EtOAc (50 mL). The EtOAcsolution was washed with sat. NaHCO₃ solution (50 mL×2), sat. NaClsolution (50 mL), dried (Na₂ SO₄), and evaporated. The residue waschromatographed on a silica column (1.9×38 cm, eluted with hexane, 10%,15% EtOAc/hexane). Evaporation of fractions 30-37 (15 mL per fraction)gave 0.593 g (45%) of 91a as a colorless syrup. MS: (El) m/e 658.0780(2%, M⁺ =658.0774). ¹ H NMR (DMSO-d₆): d 7.93 (s, 1, 7-H), 7.35, 6.92 (2x m, 15, 3×PhCH₂), 5.99 (d, 1, 1'-H, J_(1'-2') 8.0 Hz), 4.72, 4.58, 4.35(3 x m, 9, 2'-H, 3'-H, 4'-H, end 3×PhCH₂), 3.76 (dd, 1, 5'-H, J_(4'-5')=2.0 Hz, J_(5'-5'-5") 11.0 Hz), 3.65 (dd, 1,5--H, J_(4'-5") =2.5 Hz). ¹³C NMR (DMSO-d₆): d 142.00 (C2), 138.40 (C3a), 137.81, 137.44, 136.74(3×PhCH₂), 131.34 (C7a), 128.08, 128.08, 127.59, 127.40, 127.32, 127.28,126.97 (3×PhCH₂ and C4), 124.64, 122.52 (C5 and C6), 112.65 (C7), 87.45(C1'), 83.03, 77.08, 74.92 (C2', C3', and C4'), 72.43, 71.44, 7128(3×PhCH₂), 69.22 (C5').

Evaporation of fractions 44-68 (15 mL per fraction) gave 0.69 g of theα-anomer as a syrup. This sample contained a small amount of thestarting material (13) and was further purified on a silica column(1.9×16 cm, eluted with CHCl₃, 0.5% MeOH/CHCl₃). Evaporation offractions 5-10 (15 mL per fraction) gave 0.619 g (47%) of the α-anomeras a syrup. MS: (El) m/e 658.0781 (1%, M⁺ =658.0774). ¹ H NMR (DMSO-d₆):d 7.82 (s, 1,7-H), 7.32, 7.15, 6.77 (3×m, 15, 3×PhCH₂), 6.42 (d, 1,1'-H, J_(1'-2') =45 Hz), 4.58, 4.34, 4.13 (m, t, d, 9, 2'-H, 3'-H, 4'-H,and 3×PhCH₂); 3.68 (m, 2, 5'-H and 5"-H, J_(5'-5") =11.0 Hz). ¹³ C NMR(DMSO-d₆): d 141.18 (C2), 138.16, 137.89, 137.43, 136.81 (C3a and3×PhCH₂), 133.19 (C7a), 127.99, 127.93, 127.56, 127.49, 127.38, 127.22,126;98, 126.38 (3×PhCH₂ and C4), 123.91, 121.91 (C5 and C6), 113.95(C7), 87.56 (C1'), 80.77, 77.60, 77.31 (C2', C3', and C4'), 72.77,71.36, 71.82 (3×PhCH₂), 69.50 (C5').

1-(β-O-Ribofuranosyl)-2,4,5,6-tetrachlorobenzimidazole (92)

To a solution of 0.565 g (0.858 mmol) of 91a in 8 mL of CH₂ Cl₂, wasadded dropwise 8.56 mL of 1M BCl₃ /CH₂ Cl₂ at -78° C. The reactionmixture was stirred at -78° C. for 2 hr and then at -40° C. for 2 hr.MeOH (2.5 mL) was added dropwise and stirring was continued at -40° C.for 10 min. The reaction mixture was diluted with EtOAc (75 mL). TheEtOAc solution was washed with H₂ O (50 mL), sat. NaHCO₃ solution (50mL), sat. NaCl solution (50 mL), dried (Na₂ SO₄), and evaporated. Theresidue was suspended in a small amount of CHCl₃ and then filtered. Thesolid product was washed with portions of CHCl₃ and recrystallized fromMeOH to give 0.271 g (81%) of 92 as white crystals. MP: 172°-175° C.(dec). ¹ H NMR (DMSO-d₆): d 8.62 (s, 1,7-H), 5.89 (d, 1,1 '-H, J_(1'-2')=8.0 Hz), 5.52 (d, 1,2'-OH, J_(2'-2'OH) =6.5 Hz), 5.46 (t, 1, 5'-OH,J_(5'-5'OH) =4.5 Hz), 5.31 (d, 1, 3'-OH, J_(3'-3'OH) =4.5 Hz), 4.39 (m,1,2'-H, J_(2'-3') =5.5 Hz), 4.14 (m, 1,3'-H, J_(3'-4') =2.0 Hz), 4.04(m, 1, 4'-H, J_(4'-5') =J_(4'-5") =2.5 Hz), 3.72 (m, 2, 5'-H and 5"-H,J_(5'-5") =12.0 Hz). ¹³ C NMR (DMSO-d₆): d 142.44 (C2), 138.56 (C3a),131.95 (C7a), 126.91 (C4), 124.52, 122.39 (C5 and C6), 113.56 (C7),89.56 (C1'), 86,44 (C4'), 71.95 (C2'), 69.53 (C3'), 60.82 (C5"). Anal.Calcd. for C₁₂ H₁₀ Cl₄ N₂ O₄ : C, 37.14; H, 2.60; N, 7.22. Found: C,37.01; H, 2.60; N, 7.01

2-Chloro-1-(2,3,5-tri-O-acetyl-β-D-ribofuranosyl)-4,5,6-tribromobenzimidazole(102)

To a suspension of 0.779 g (2 mmol) of 20 in 10 mL of dry MeCN, wasadded 0.5 mL (2 mmol) of BSA. The reaction mixture was stirred at 75° C.for 15 min. This solution was treated with 0.700 g (2.2 mmol) of1,2,3,5-tetra-O-acatyl-β-D-ribofuranose and 0.43 mL (2.2 mmol) of TMSOTfat 75° C. for 1 hr. The reaction mixture was cooled and diluted withEtOAc (50 mL). The EtOAc solution was washed with sat. NaHCO₃ solution(50 mL×2), sat. NaCl solution (50 mL), dried (Na₂ SO₄), and evaporated.The residue was recrystallized from MeOH to give 1.1 g of the blockednucleoside as white crystals. This product contained some Impurity byTLC and was further purified on a silica column (1.9×25 cm, eluted withCHCl₃). Evaporation of fractions 8-20 (20 mL per fraction) andrecrystallization from MeOH gave 0.974 g (2 crops, 75%) of 102 as whitecrystalline needles. MP: 163°-165° C. MS: (El) m/e 643.8201 (2%, M⁺=643.8196). ¹ H NMR (DMSO-d₆): d 8.25 (5, 1,7-H), 6.25 (d, 1, 1'-H,J_(1'-2') =7.0 Hz), 5.53 (t, 1, 2'-H, J_(2'-3') =7.0 Hz), 5.43 (dd, 1,3'-H, J_(3'-4') =4.5 Hz), 4.47 (dd, 5'-H, J_(5'-4') =5.0 Hz, J_(5'-5")=11.5 Hz), 4.46 (m, 1, 4'-H), 4.37 (dd, 1, 5"-H, J_(5'-4') =1.5 Hz),2.15, 2.13, 2.01 (3 x s, 9, 3 x Ac). ¹³ C NMR (DMSO-d₆): d 169.93,169.42, 169.16 (3×COCH₃), 140.70 (C2), 140.85 (C3a), 132.58 (C7a),121.30 (C5), 119.07 (C6), 115.93 (C4), 115.72 (C7), 86.94 (C1'), 70.73(C2'), 68.62 (C3'), 62.53 (C5'), 20.66, 20.26, 19.97 (3×COCH₃). Anal.Calcd. for C₁₈ H₁₆ Br₃ ClN₂ O₇ : C, 33.39; H, 2.49; N, 4.33. Found: C,33.30; H, 2.33; N 4.31.

2-Chloro-1-(β-D-ribofuranosyl)-4,5,6-tribromobenzimidazole (103)

A mixture of 0.430 g (0.664 mmol) of the blocked nucleoside (102) and0.216 g (3.317 mmol) of KCN in 13 mL of 70% EtOH was stirred at roomtemperature for 6 hr. The reaction mixture was diluted with 75 mL ofEtOAc. The EtOAc solution was washed with H₂ O (50 mL), sat. NaClsolution (50 mL), dried (Na₂ SO₄), and evaporated. The residue waschromatographed on a silica column (1.9×15 cm, eluted successively withCHCl₃, 1%, 2%, 3% MeOH/CHCl₃). Evaporation of fractions 43-60 (10 mL perfraction) and recrystallization from MeOH/MeCN gave 0.157 g (2 crops,44%) of 103 as white crystals. MP: 168°-170° C. MS: (FAB) m/e 518.7947(28%, MH⁺ =518.7957). ¹ H NMR (DMSO-d₆): d 8.76 (s, 1,7-H), 5.87 (d, 1,1 '-H, J_(1'-2') =8.0 Hz), 5.51 (d, 1,2'-OH, J_(2'-2'OH) =6.0 Hz), 5.42(t, 1, 5'-OH, J_(5'-5'OH) =4.5 Hz), 5.30 (d, 1, 3'-OH, J_(3'-3'OH) =4.5Hz), 4.37 (m, 1, 2'-H, J_(2'-3') =5.5 Hz), 4.14 (m, 1,3'-H, J_(3'-4')=2.0 Hz), 4.02 (m, 1,4'-H, J_(4'-5') =J_(4'-5") =2.5 Hz), 3.71 (m, 2,5'-H and 5"-H, J_(5'-5") =12.0 Hz). ¹³ C NMR (DMSO-d_(6l) ): d 142.32(C2), 140.94 (C3a), 132.50 (C7a), 120.62 (C5), 118.44 (C6), 117.10 (C7),115.34 (C4), 89.58 (C1'), 86.50 (C4'), 71.93 (C2'), 69.63 (C3'), 60.89(C5'). Anal. Calcd. for C₁₂ H₁₀ Br₃ ClN₂ O₄ 0.3 MeCN: C, 28.36; H, 2.06;N, 6.04. Found: C, 28.25; H, 1.84; N, 5.90. (the Content of 0.3 MeCN wasalso indicated by ¹ H NMR.)

2-Azido-5,6-dichloro-1-(2-deoxy-3,5-di-O-p-toluoyl-β-D-erythro-pentofuranosyl)benzimidazole(110a)

Compound 110 (5.74 g, 10 mmol) was stirred with 4.90 g (100 mmol) ofLiN₃ in 50 mL of dry DMF at 70° C. for 16 hr. The reaction mixture wasevaporated and coevaporated with toluene. The residue was partitionedbetween EtOAc/half sat. NaCl solution (250 mL/200 mL). The EtOAc layerwas washed with half sat. NaCl solution (200 mL), dried (Na₂ SO₄),evaporated. The residue was dried under oil-pump vacuum at roomtemperature for 2 hr to give 110a as a yellowish foam (6.0 g,˜103%).This sample was pure by TLC and was used directly in the subsequentreactions without further purification. Analytical sample was obtainedby recrystallization from EtOH. MP: 82°-85° C. MS: (El) m/e 579.1065(2.5%, M⁺ =579.1076). ¹ H NMR (DMSO-d₆): d 7.97, 7.81 (2 x s, 2, 7-H and4-H), 7.96, 7.88, 7.36, 7.30 (4 x d, 8, Ph, J=8.0 Hz), 6.36 (dd, 1,1'-H, J_(1'-2') =8.5 Hz, J_(1'-2') =6.0 Hz), 5.72 (m, 1, 3'-H, J_(3'-2')=7.0 Hz, J_(3'-2") =2.0 Hz, J_(3'-4') =3.35 Hz), 4.71 (dd, 1, 5'-H,J_(5'-4') =3.5 Hz, J_(5'-5") =12.0 Hz), 4.63 (dd, 1,5"-H, J_(5'-4') =5.0Hz), 4.53 (m, 1, 4'-H), 3.05 (m, 1, 2'-H, J_(2'-2") =15.0 Hz), 2.60 (m,1, 2"-H), 2.40, 2.36 (2 x s, 6, 2 x Me). ¹³ C NMR (DMSO-d₆): d 165.46,165.31 (2×p-MePhCO), 149.07 (C2), 144.04, 143.78 (2×p-MePhCO), 140.52(C3a), 132.63 (C7a), 129.49, 129.24 129.20 (2 x p-MePhCO), 126.55,126.43, 125.35, 124.57 (2×p-MePhCO, C6, and C5), 118.98 (C4), 112.78.(C7), 63.78 (C1'), 81.19 (C4'), 73.90 (C3'), 63.72 (C5'), 35.31 (C2'),21.15, 21.10 (2×p-MePhCO). Anal. Calcd. for C₂₈ H₂₃ Cl₂ N₅ O₅ : C,57.94; H, 3.99; N, 12.07. Found: C, 58.07; H, 4.12; N, 11.94.

2-Azido-5,6-dichloro-1-(2-deoxy-β-D-erythro-pentofuranosyl)benzimidazole(110b)

Compound 110a (6.0 g, prepared from 10 mmol of 110) was treated with 150mL of NH₃ /MeOH at room temperature for 2 days. The reaction mixture wasevaporated and coevaporated with MeOH. The resulting solid wastriturated with CHCl₃ (50 mL). The CHCl₃ suspension was kept at 0° C.for 1 hr and was then filtered. The solid was washed with portions ofCHCl₃ and then recrystallized from MeOH to give 2.462 g (2 crops) of110b as white needles. The CHCl₃ filtrate was concentrated to 20 mL andkept in a freezer (˜-15° C.) overnight. The precipitate was collectedand recrystallized from MeOH to give additional 0.569 g (2 crops) of110b. The total yield was 3.031 g (88% from 110); MP: 159°-161° C.(dec). MS: (El) m/e 343.0232 (30%, M⁺ =343.0239). ¹ H NMR (DMSO-d₆): d8.26 (s, 1,7-H), 7.81 (s, 1, 4-H), 6.09 (dd, 1, 1 '-H, J_(1'-2') =9.0Hz, J_(1'-2") =6.0 Hz), 5.36 (d, 1, 3'-OH, J_(3'-3'OH) =4.5 Hz), 5.15(t, 1, 5'-OH, J_(5'-5'OH) =5.0 Hz), 4.39 (m, 1,3'-H, J_(3'-2') =6.5 Hz,J_(3'-2') =2.5 Hz, J_(3'-4') =3.0 Hz), 3.84 (m, 1,4'-H, J_(4'-5') =3.5Hz), 3.65 (m, 2,5'-H), 250 (m, 1, 2'-H, J_(2'-2') =13.0 Hz), 2.09 (m, 1,2"-H). ¹³ C NMR (DMSO-d_(6l) ): d 149.00 (C2), 140.90 (C3a), 132.11(C7a), 125.13, 124.25 (C5 and C6), 118.79 (C4), 114.11 (C7), 87.52(C4'), 84.08 (C1'), 70.15 (C3'); 60.10 (5'), 38.54 (C2'). Anal. Calcd.for C₁₂ H₁₁ Cl₂ N₅ O₃ : C, 41.88; H, 3.22; N, 20.38. Found: C, 42.01; H,3.13; N, 19.98.

2-Amino-5,6-dichloro-1-(2-deoxy-β-D-erythro-pentofuranosyl)benzimidazole(113)

A mixture of 110b (1.032 g, 3 mmol) and Ra-Nl (0.24 g, wet) in 30 mL ofEtOH was hydrogenated (50 psi of H₂) at room temperature for 6 hr. Thereaction mixture was filtered and the filtrate was evaporated. Theresidue was suspended in 20 mL of MeCN and the suspension was filteredto give 0.856 g (90%, 2 crops) of 113 as a white solid. Analyticalsample (as white needles) was obtained by recrystallization fromMeOH/MeCN. MP: 216°-221° C. (dec.). MS: (El) m/e 317.0340 (37% M⁺=317.0334). ¹ H NMR (DMSO-d₆): d 7.71 (s, 1, 7-H), 7.29 (s, 1, 4-H),6.93 (br s, 2, 2-NH₂), 6.23 (dd, 1, 1 '-H, J_(1'-2') =8.5 Hz, J_(1'-2')=6.0 Hz), 5.40 (t, 1, 5'-OH, J_(5'-5'OH) =4.5 Hz), 5.34 (d, 1, 3'-OH,J_(3'-3') OH=4.0 Hz), 4.40 (m, 1, 3'-H, J_(3'-2') =7.0 Hz, J_(3'-2")=2.0 Hz, J_(3'-4') =2.5 Hz), 3.82 (m, 1, 4'-H, J_(4'-5') =2.5 Hz), 3.68(m, 2, 5'-H), 2.40 (m, 1, 2'-H, J_(2'-2') =13.0 Hz), 2.05 (m, 1, 2"-H).¹³ C NMR (DMSO-d₆): d 155.65 (C2), 143.16 (C3a), 132.40 (C7a), 122.94,119.77 (C5 and C6), 115.35 (C4), 110.69 (C7), 86.91 (C4'), 83.87 (C1'),70.20 (C3'), 60.74 (C2'). Anal. Calcd. for C₁₂ H₁₃ Cl₂ N₃ O₃ : C, 45.30;H, 4.12; N, 13.21. Found: C, 45.44; H, 4.10; N, 13.28.

2,5,6-Trichloro-1-(2,3,5-tri-O-benzyl-β-D-arabinofuranosyl)benzimidazole(133)

Compound 5 (598 mg 2.7 mmole) was dissolved in dry CH₃ CN(60 ml) and 97%NaH (80 mg, 3 mmole) was added. The mixture was stirred under N₂ for 1hour at room temperature. Then 2,3,5-tri-O-benzyl-D-arabinofuranosylchloride (prepared from 2,3,5-tri-O-benzyl-1-O-p-nitrobenzoyl-D-arabinofuranose) (1.83 g, 3 mmole)) dissolved in dryCH₃ CN (20 ml) was added dropwise. The mixture was stirred under N₂ forovernight at room temperature. The reaction solution was filteredthrough Celite. The filtrate was evaporated to give a syrup and thissyrup was subjected to flash column chromatography on silica gel(Kieselgel 60, 230-400 mesh) and eluted with CH₂ Cl₂. The fractionscontaining 133 were combined and evaporated to give 1.6 g (95.0%) of133. (IR KBr cm⁻¹ : 1140-1060(C--O--C)).

2,5,6-Trichloro-1-(β-D-arabinofuranosyl)benzimidazole (134)

Compound 133 (1.25 g 2 mmole) was dissolved in dry CH₂ Cl₂ (20 ml) andthis solution was cooled in an acetone-dry ice bath. A 1M solution ofBCl₃ in CH₂ Cl₂ (10 ml) was added and the mixture was stirred at -70°for 4.5 hr. At this point TLC showed no starting material. CH₃ OH (10ml) was added and the solution was made neutral with NH₄ OH. Thesolution was evaporated in vacuo to dryness. H₂ O (20 ml) was added andthe solution was extracted with EtOAc. The EtOAc layer was washed withH₂ O, dried (Na₂ SO₄) and evaporated in vacuo to obtain a syrup. Thissyrup was subjected to flash column chromatography on silica gel(Kieselgel 60, 230-400 mesh) with CH₂ Cl-MeOH (20:1) as eluent solventto give 134. Yield 0.551 g, 77.9%.

2,4,5,6,7-Pentachloro-1- (1,3-dibenzyloxy-2-propoxy)methyl!benzimidazole(154)

The title compound was prepared from 2,4,5,6,7-pentachlorobanzimidazole(14) (0.29 g, 1.0 mmole), CH₃ CN (35 mL), NaH (97%, oil dispersion,0.033 g, 1.4 mmole) and (1,3-bis(banzyloxy)-2-propoxy!methylchloride(0.49 g, 1.5 mmole ) in CH₃ CN (10 mL) to afford an oil. The resultingoil was purified by flash column chromatography (2×20 cm) (230-400 mesh)prepared with wet SiO₂ in hexane. Elution with hexane:EtOAc (8:2,v/v)gave 0.32 g of the blocked product 134 as an oil. Although the ¹ H NMRspectra revealed a small amount of an impurity, the compound was usedwithout further purification for the next step.

2,4,5,6,7-Pentachloro-1- (1,3-dihydroxy-2-propoxy)methyl!benzimidazole(155)

Compound 155 (0.52 g, 51%) was prepared from 154 (1.5 g, 2.6 mmole),BCl₃ (5M solution in CH₂ Cl₂, 14 mL), CH₂ Cl₂ (30 mL) and MeOH (20 mL)by the method described for 166 which on recrystallization from EtOACgave 155: m.p. 155°; ¹ H NMR (DMSO-d₆): d 5.76 (s, 2 H, C-1'), 4.63 (t,2 H, exchanges with D₂ O, 2×OH), 3.54-3.48 (m, 1 H, C-3'), 3.43-3.25 (m,4 H, C-4' and 5'H). Anal. Calcd. for C₁₁ H₉ N₂ Cl₅ O₃ : C, 33.46; H,2.28; N, 7.09. Found: C, 33.60; H, 2;15; N, 6.91.

2-Methoxy-4,5,6,7-tetrachloro-1-(1,3-dihydroxy-2-propoxy)methyl!benzimidazole (157a) and2-Amino-4,5,6,7-tetrachloro-1- (1,3-Dihydroxy-2-propoxy)methyl!benzimidazole (156)

A mixture of 155 (0.28 g, 0.71 mmole) and methanolic ammonia (methanolsaturated with ammonia at 0° C. 60 mL) was stirred at room temperaturefor 36 hr in a pressure bottle. The resulting mixture was evaporated todryness and was purified by flash column chromatography (3×15 cm)(230-400 mesh). Elution of the column CH₂ Cl₂ :MeOH (97:3, v/v) andevaporation of the appropriate fractions followed by recrystallizationfrom of MeOH gave 0.06 gm (20 %) of 157a: mp. 192° C.; ¹ H NMR(DMSO-d₆): d 5.71 (s, 2 H, C-1'), 4.59 (t, 2 H, exchanges with D₂ O, 2 xOH ), 4.19 (s, 3H, OCH₃), 3.75-3.21 (m, 5 H, C-3', C-4' and C-S'). Anal.Calcd. for C₁₂ H₁₂ N₂ Cl₄ O₄ : C, 36.92; H, 3.07; N, 7.17. Found: C,37.20; H, 2.99; N, 6.93,

Further elution of the CH₂ Cl_(2:) MeOH (90:1, v/v) and evaporation ofthe appropriate fractions followed by recrystallization from the mixtureof DMSO-H₂ O gave 0.2 g (69%) of 156: m.p. 268° C.; ¹ H NMR (DMSO-d₆): d7.40 (bs, 2 H, exchanges with D₂ O, NH₂), 5.75 (s, 2 H, C1'), 4.77 (156,1 H, exchanges with D₂ O, 2 x OH ), 3.58-3.34 (m, 5 H, C-3', C-4' andC-S'). Anal. Calcd. for C₁₁ H₁₁ N₃ Cl₄ O₃ : C, 35.20; H, 2.93; N, 11.20.Found: C, 35.37; H, 3.02; N, 10.86.

2,4,5,6,7-Pentachloro-1-(2-acetoxyethoxymethyl)benzimidazole (165)

NaH (0.12 g, 3.0 mmole, 60% oil disperson) was added to a stirredsuspension of 2,4,5,6,7-pentachlorobenzimidazole (14) (1.0 g, 3.4.mmole)in dry CH₃ CN (120 mL) under a N₂ atmosphere. The solution was surfeduntil H₂ evolution has ceased and a clear solution was obtained (20min.). (2-Acetoxyethoxy) methyl bromide (0.68 g, 10.68 mmole) in CH₃ CN(34 mL) was then added dropwise. The reaction mixture was stirred for anadditional 3 hr. The resulting mixture was concentrated under reducedpressure, diluted with H₂ O (50 mL), extracted with EtOAc (100 mL),washed with H₂ O (100 mL), dried (anhyd. Na₂ SO₄) and the solvent wasremoved under reduced pressure to yield an oil (165). The resulting oilwas purified by flash column chromatography (2×24 cm) (230-400 mesh).Elution of the column with hexane:EtOAc (6:4, v/v) and evaporation ofthe appropriate fractions gave a solid which was recrystallized fromMeOH to afford 1.39 g (72%) of 165: m.p. 119° C. ¹ H NMR (CDCl₃): d 5.93(s, 2 H, C-1'), 4.143.92 (m, 4 H, C-3' and C-4'), 1.83 (s, 3 H, Ac).Anal. Calcd. for C₁₂ H₉ N₂ Cl₅ O₃ : C,35.45; H, 2.23; N, 6.89. Found: C,35.57; H, 2.17; N, 16.91.

2,4,5,6,7-Pentachloro-1 -(2-benzyloxyethoxymethyl)benzimidazole

NaH (0.033 g, 1.52 mmole, 90% oil disperson) was added to a stirredsuspension of 2,4,5,6,7-pentachlorobenzimidazole (14) (0.3 g, 1.03mmole) in dry CH₃ CN (35 mL) under a N₂ atmosphere. The Solution wasstirred until H₂ evolution had ceased and a clear solution was obtained(20 min.). (2-Benzyloxyethoxy)methylchloride (0.31 g, 1.54 mmole) in CH₃CN (10 mL) was then added dropwise. The reaction mixture was stirred foran additional 15 hr. The resulting mixture was concentrated underreduced pressure, diluted with H₂ O (50 mL), extracted with EtOAc (100mL), washed with H₂ O (100 mL), dried (anhyd. Na₂ SO₄) and the solventwas removed under reduced pressure to yield an oil. The resulting oilwas purified by flash column chromatography (2×24 cm) (230-400 mesh).Elution of the column with hexane:EtOAc (6:4, v/v) and evaporation ofthe appropriate fractions followed by crystallization from hexane:EtOAcafforded 0.18 g (38.4%) of blocked product: m.p. 95° C.; ¹ H NMR(CDCl₃): d 7.22-7.34 (m, 5H, C₆ H₅), 5.93 (s, 2 H, C-1'), 4.47 (s, 2H,CH₂ C₆ H₅), 3.70-3.73 (m, 2 H, C-3'), 3.45-3.57 (m, 2 H, C-4'), 1.83 (s,3 H, Ac). Anal. Calcd. for C₁₇ H₁₃ NCl₅ O₂ : C, 44.88; H, 2.86; N, 6.16.Found: C, 44.95; H, 2.59; N, 6.05.

2,4,5,6,7-Pentachloro-1-(2-hydroxyethoxymethyl)benzimidazole (166)

To a solution of the blocked product (1.2 g, 2.63 mmole) in dry CH₂ Cl₂(25 mL) at -78° C. under N₂ atmosphere, BCl₃ (5M solution in CH₂ Cl₂, 18mL) was added dropwise while maintaining the bath temperature at 78° C.The reaction mixture was stirred for an additional 4 hr, MeOH (18 mL)was added at 0° C. and the cold solution was immediately neutralized (pH7) with NH₄ OH. The solution was allowed to warm up to room temperatureand then concentrated at 40° C. to yield an oil. The resulting mixturewas diluted with H₂ O (100 mL), extracted with EtOAc (250 mL), washedwith H₂ O (100 mL), dried (anhyd. Na₂ SO₄) and concentrated in vacuo.The product thus obtained, was purified by flash column chromatography(2×24 cm) (230-400 mesh). Elution of the column with hexane:EtOAc (6:4,v/v) and evaporation of the appropriate fractions gave a solid which onrecrystallization from hexane:EtOAc afforded 0.45 gm (45%) of 166: m.p.168°-170° C.; ¹ H NMR (DMSO-d₆): d 5.83 (s, 2H, C-1'), 4.62 (t, 1 H,exchanges with D₂ O, OH), 3.54-3.50 (m, 2 H, C-3'), 3.44-3.38 (m, 2 H,C-4'). Anal. Calcd. for C₁₀ H₇ N₂ Cl₅ O₂ : C, 32.92; H, 1.92; N, 7.68.Found: C, 32.58; H, 1.82; N, 7.33.

2-Methoxy-4,5,6,7-tetrachloro-1-(2-hydroxyethoxymethyl)benzimidazole(166a) and2-Amino-4,5,6,7-tetrachloro-1-(2-hydroxyethoxymethyl)benzimidazole (167)

A mixture of 165 (0.2, 0.49 mmole) and methanolic ammonia (methanolsaturated with ammonia at 0° C., 30 mL) was stirred at room temperaturefor 12 hr in a pressure bottle. The resulting mixture was evaporated todryness and purified by flash column chromatography (2×25 cm) (230-400mesh). Elution of the column CH₂ Cl₂ : MeOH (97:3, v/v) and evaporationof the appropriate fractions followed by recrystallization from themixture of hexane:EtOAc gave 0.02 gm (13.24%) of 166a: m.p. 139° C.; ¹ HNMR (DMSO-d₆): d 5.44 (s, 2 H, C-1'), 4.69 (t, I H, exchanges with D₂ O,OH), 3.86 (s, 3 H, OCH₃), 3.46-3.41 (m, 4 H, C-3' and C-4'). Anal.Calcd. for C₁₁ H₁₀ N₂ Cl₄ O₂ : C; 34.81; H, 3.62; N, 12.16. Found: C,34.59; H, 2.55; N, 11.79.

Further elution of the CH₂ Cl₂ : MeOH (90:1, v/v) and evaporation of theappropriate fractions followed by recrystallization from the mixture ofDMSO-H₂ O gave 0.06 g (39.5%) of 167: m.p. 268° C.; ¹ H NMR (DMSO-d₆): d7.42 (s, 2 H, exchanges with D₂ O, NH₂), 5.66 (s, 2 H, C-1'), 4.69 (bs,1 H, exchanges with D₂ O, OH ), 3.47 (m, 4 H, C-3' and C-4'). Anal.Calcd. for C₁₀ H₉ N₃ Cl₄ O₂ : C, 36.62; H, 2.79; N, 7.78. Found: C,36.61; H, 2.16; N, 7.65.

1 -Benzyl-2,5,6-trichlorobenzimidazole (181)

2,5,6-Trichlorobenzimidazole (5, 50 mg, 0.0002 moles) was dissolved in15 ml of CH₃ CN, and K₂ CO₃ (33 mg, 0.0002 moles) was added. The mixturewas stirred at room temperature for one hour, at which time benzylbromide (0.024 ml, 0.0002 moles) was added. The mixture was then stirredfor two days. TLC analysis showed the formation of a new compound with ahigher R_(f) than starting material. The mixture was then concentratedand a liquid-liquid extraction was done using CHCl₃, and H₂ O. The CHCl₃layer was kept, dried with MgSO₄, filtered, and then concentrated.Column chromotograpy was then performed using 2% MeOH/CHCl₃ to elute thecompounds. The fractions were collected and those containing thecompound were concentrated. The compound was recrystallized from MeOH,giving 50 mg of 1-benzyl-2,5,6-trichlorobenzimidazole (181) (96.2%). ¹ HNMR (CDCl₃): d 3.49ppm (s, 3H), 7.13 (m, 2H), 7.32 (m, 4H), 7.78 (s,1H). ¹³ C NMR (CDCl₃): d 43.20 ppm, 106.25, 115.78, 121.62, 122.24,122.66, 123.50, 124.15, 129.05, 129.16, 135.97, 137.68. GC-MS: m/e 310,273, 239, 221,197, 170, 158, 138, 120, 100,91, 77, 65, 51, 39. m.p:175°-176° C. Elemental Analysis: (C₁₄ H₉ Cl₃ N₂): Calculated: C, 53.85;H, 2.88; N, 8.97. Found: C, 53.92; H, 2.85; N, 8.89. UV Imax nm (e×104):(pH 7) 263 (0.080), 290 (0.102), 299 (0.154); (pH 1), 224 (0.412), 258(0.157),290 (0.155), 299 (0.168).

2-Amino-1-benzyl-5,6-dichlorobenzimidazole (182)

Method A(182)

2-Amino-5,6-dichlorobenzimidazole (4) (1 g, 4.95 mmole) was dissolved inacetonitrile (200 ml) and NaOH (198 mg, 4.95 mmole) was added. Afterstirring for 1 hr at room temperature, benzyl bromide (0.589 ml, 4.95mmole) was added and the mixture was allowed to stir overnight. Themixture was concentrated under reduced pressure and purified on a silicagel column (3.5×5 cm) using 5% MeOH/CHCl₃ to give 1.23 g (85%) of2-amino-1-benzyl-5,6-dichlorobenzimidazole (182). The compound was thenrecrystallized from EtOH/H₂ O to give a white powder. ¹ H NMR (DMSO-d₆):d 5.28 ppm (s, 2H), 6.95 (s, 2H), 7.16 (d, 2H), 7.25 (d, 1H), 7.31 (m,4H). ¹³ C NMR (DMSO-d₆); d 44.74 ppm, 109.15, 115.37, 119.62, 122.66,125.82, 127.37, 128.55, 134.10, 136.46, 143.13, 156.91. MS (ElectronImpact): m/e 291, 200, 158, 91, 65. MP: 245°-246° C. Elemental Analysis:(C₁₄ H₁₁ Cl₂ N_(3')): Calculated: C, 57.55; H, 3.79; N, 14.58. Found: C,57.66; H, 3.83; N, 13.98. UV Imax nm (e×104): (pH 7) 230 (0.761), 260(0.324), 302 (0.452); (pH 1) 238 (0.306), 294 (0.383), 299 (0.359); (pH11) 229 (0.869), 257 (0.316) 301 (0.443).

Method B(182)

Compound 118 was treated with methanolic ammonia for 18 hr, at anelevated temperature, to afford compound 182, after a standard isolationand purification procedure.

D. Antiviral Activity of Compounds

The following test methods were followed in generating the data inTables 1 through 3 and FIGS. 3 through 6.

METHODS

(1) Propagation of Cells and Viruses

(a) Cells

The routine growth and passage of KB cells, a human epidermoidneoplastic cell line, was performed in monolayer cultures using minimalessential medium (MEM) with either Hanks Salts MEM(H)! or Earle salts (MEM(E)! supplemented with 10% calf serum or 5 to 10% fetal bovine serum.The sodium bicarbonate concentration was varied to meet the bufferingcapacity required. BSC-1 (African green monkey kidney) cells were grownand passaged in Dulbecco modified MEM(E) supplemented with 5% tryptosephosphate broth and 5% horse serum. Cultures of human foreskinfibroblasts (HFF) were grown in medium consisting of MEM(E) with 10%fetal bovine serum.

Cells were passaged at 1:2 to 1:10 dilutions according to conventionalprocedures by using 0.05% trypsin plus 0.02% EDTA in a HEPES bufferedsalt solution. HFF cells were passaged only at 1:2 dilutions.

(b) Viruses

The S-148 strain of HSV-1 was used in most experiments and was providedby Dr. T. W. Schafer of Schering Corporation. The HF strain of HSV-1 wasused in selected experiments and was obtained from Dr. G. H. Cohen,University of Pennsylvania. The Towne strain, plaque-purified isolateP_(o') of HCMV was a gift of Dr. Mark Stinski, University of Iowa.

High titer HSV-1 stocks were prepared as follows: Nearly confluentmonolayer cultures of KB cells were grown in 32 oz. glass bottlescontaining MEM(E) buffered with 25 mM HEPES and supplemented with 5%fetal bovine serum and 0.127 g/liter L-arginine (VGM, virus growthmedium). The cultures were infected at a low input multiplicity toreduce the formation of defective virus. After cell cytopathologyreached "three to four plus", the cells were harvested by vigorousshaking, and concentrated by centrifugation (800 x g for 5 min.). Theresulting virus pools were stored at -76° C. until retrieved for use inexperiments.

Stock HCMV was prepared by infecting HFF cells at a multiplicity ofinfection (m.o.i.) of less that 0.01 plaque-forming units (p.f.u.) percell. Cell growth medium was changed every four days until cytopathologywas evident in all cells (approximately 21 days). Supernatant fluidswere retained as the virus stock. Four days later, the remaining cellswere disrupted by three cycles of freeze-thawing and the cell plusmedium held as an additional source of virus. Storage was in liquidnitrogen.

HSV-1 was titered using monolayer cultures of BSC-1 cells. Cells wereplanted at 3×10⁵ cells/well using 6-well cluster dishes. MEM(E)supplemented with 10% fetal bovine serum was employed as medium. After22-24 h, cells were 90% confluent and were inoculated in triplicateusing at least three ten-fold dilutions with 0.2 ml of the virussuspension to be assayed and incubated in a humidified 4% CO₂ -90% airatmosphere for one hour ;to permit viral adsorption. Following virusadsorption, the cell sheet was overlayed with 5 ml of MEM(E) with 5%serum plus 0.5% methocel (4000 CPS) and incubated an additional two tothree days. Cells were fixed and stained with 0.1% crystal violet in 20%methanol and macroscopic plaques enumerated.

HCMV was titered in 24-well cluster dishes which were planted to contain5×10⁴ HFF cells/well, grown as described above. When the cells were 70to 80% confluent, 0.2 ml of the virus suspension was added per well andadsorbed as described above. At least three ten-fold dilutions of eachpreparation were used. Following virus adsorption, the cell sheets wereoverlayed with 0.5% methocel (4000 CPS) in maintenance medium MEM(E)with 1.1 g/liter NaHCO₃, 100 units/ml penicillin G, 100 μg/mlstreptomycin, and 5% fetal bovine serum!. The cultures were incubated ina humidified atmosphere of 4% CO₂ -96% air. Viral foci were visible 5 to7 days after infection using at least 10-fold magnification. Cells werefixed and stained by a 10-minute exposure to a 0.1% solution of crystalviolet in 20% methanol 7 to 12 days after infection. Microscopic fociwere enumerated at 20-fold magnification using a Nikon ProfileProjector.

(2) Assays for Antiviral Activity

(a) HSV-1

Plaque reduction experiments with HSV-1 were performed using monolayercultures of BSC-1 cells. The assay was performed exactly as describedabove except that the 0.2 ml virus suspension contained approximately100 p.f.u. of HSV-1. Compounds to be tested were dissolved in theoverlay medium at concentrations usually ranging from 0.1 to 100 μM inhalf-or-one logarithm₁₀ dilutions. Titer reduction assays were performedby planting KB cells in 25 cm² plastic tissue culture flasks 10 to 24 hrprior to infection. At the onset of experiments, logarithmically growingreplicate monolayer cultures were 60 to 80% confluent and contained 2.5to 4.5×10⁶ cells/flask. Medium was decanted and the cultures wereinfected with 2 to 10 p.f.u. of HSV-1 per cell. Virus was contained in1.0 ml of VGM supplemented with 5% fetal bovine serum. After a 1 hadsorption period at 37° C., the cell sheet was rinsed twice with 2 mlof VGM without serum to remove unadsorbed virus and 5 ml of VGMcontaining drugs at three to five selected concentrations added induplicate. Following an 18-to 20-hr incubation at 37° C., infectedmonolayers were treated with EDTA-trypsin to suspend the cells; aliquotswere removed, subjected to three cycles of freezing and thawing, andstored at -76° C. for subsequent virus assay. Virus was titered on BSC-1cells as described above.

ELISA techniques according to standard procedures were also used todetermine activity against HSV-1. Drug effects were calculated as apercentage of the reduction in virus titers in the presence of each drugconcentration compared to the titer obtained in the absence of drug.Acylovir was used as a positive control in all experiments.

(b) HCMV

The effect of compounds of the replication of HCMV has been measuredusing both a plaque (focus) reduction assay and a titer (yield)reduction assay. For the former, HFF cells in 24-well culture disheswere infected with approximately 50 p.f.u. of HCMV per well using theprocedures detailed above. Compounds dissolved in growth medium wereadded in three to six selected concentrations to triplicate wellsfollowing virus adsorption. Following incubation at 37° C. for 7 to 10days, cell sheets were fixed, stained and microscopic plaques wereenumerated as described above. Drug effects were calculated as apercentage of reduction in number of foci in the presence of each drugconcentration compared to the number observed in the absence of drug.DHPG (ganciclovir) was used as a positive control in all experiments.

For titer reduction assays, HFF cells were planted as described above in24-well cluster dishes or in 25 cm² flasks. When monolayers wereapproximately 70% confluent, HCMV was added at a m.o.i. of 0.5 p.f.u.per cell and adsorbed as detailed above. Compounds dissolved in growthmedium were added in one or one-half-logarithm₁₀ dilutions andincubation continued at 37° C. After 7 to 10 days of incubation, culturedishes of flasks were frozen at 76° C. For titer determination, cellswere thawed and then subjected to two more cycles of freezing andthawing at 37° C. Serial, one-logarithm₁₀ dilutions of the finalsuspension were prepared and inoculated onto new cultures of HFF cells.Titer determination was as detailed above in part (b).

(3) Cytotoxicity Assays

(a) Protocol for Determining Effects of Compounds on DNA, RNA andProtein Synthesis

KB or HFF cells were planted using a Costar Transplate-96 (Costar,Cambridge, Mass.) in Costar 96-well cluster dishes at a concentration of10,000 to 12,000 cells per well. Cells were suspended in 200 μl ofmedium MEM(H) plus 0.7 g/liter NaHCO₃ supplemented with 10% calf serum!per well. After incubation of 16 to 24 hours at 37° C. in a humidifiedatmosphere of 4% CO₂ in air, 150 μl of medium was removed per well.One-hundred. μl of medium with or without compounds in twice their finalconcentrations was added to each well using a Titertek MultichannelPipette. Final concentrations of compounds ranged from 0.1 to 320 μl ofmedium containing radioactive precursors also was added to each well togive a final concentration to 1 to 3 μCi/ml of labeled precursor. ³H!Thd was diluted with unlabeled dThd to give a final concentration of3to 61 μM.

Following addition of drugs and labeled precursors, plates wereincubated as described above for an additional 18 to 24 hr. Logarithmiccell growth occurred during this time with continual uptake of labeledprecursors. At the end of the incubation period, cells were individuallyharvested from each well using a Skatron Cell harvester (Skatron, Inc.,Sterling, Va.). Cultures for individual wells were harvested onto filterpaper and washed free of unincorporated label with nine sequentialwashes with 5% trichloroacetic acid, nine washes with water, and ninewith ethanol using the Skatron unit. Filters were dried, circles fromindividual cultures were punched from the filter mat and placed intomini-vials. Liquid scintillation solution was added, and radioactivitydetermined in a Beckman model LS8100 Liquid scintillation spectrometer.All samples were counted for 2.0 min each, with three round of counting.Counts per minute were determined following the application ofstatistical methods to eliminate count rates which fell outsidedistribution limits defined by Chauvenetys rejection criterion.

All analyses were performed in triplicate. That is, three culture wellswere used per time point, radioactive precursor, and drug concentrationin all experiments. Results from triplicate assays were converted topercent of control and plotted as log dose-response curves from which50% inhibitor (I₅₀) concentrations were interpolated. Threeconcentrations of vidarabine were included on all plates as a positivecontrol.

(b) Visual Scoring

Cytotoxicity produced in HFF and BSC-1 cells was estimated by visualscoring of cells not affected by virus infection in the HCMV and HSV-1plaque reduction assays. Cytopathology was estimated at 35- and 60-foldmagnification and scored on a zero to four plus basis. Wells were scoredon the day of staining.

(4) Cell Growth Rates

Population doubling times and cell viability were measured in uninfectedHFF and/or KB cells. Cells were planted in replicate 6-well plastictissue culture dishes or in 25 cm² flasks as described above in part 1.Following an incubation period during which cells attached to thesubstrate, medium was decanted, the cell sheet rinsed once with HBS, andfresh medium added. The medium consisted of MEM(E) with 1.1 g NaHCO₃/liter and 10% fetal bovine or calf serum plus appropriate log orhalf-log concentrations of drug. After additional periods of incubationfrom 1 to 72 hours at 37° C., cells were harvested by means of 0.05%trypsin plus 0.02% EDTA in a HEPES-buffered salt solution. Cells wereenumerated using either a Coulter counter or a homocytometer andviability determining using trypsan blue dye exclusion.

(5) Plating Efficiency

A plating efficiency assay was used to confirm and extend resultsdescribed above. Briefly, KB cells were suspended in growth medium andan aliquot containing 1000 cells was added to a 140×25 mm petri dish.Growth medium (40 ml) containing selected concentrations of testcompounds was added and the cultures incubated in a humidifiedatmosphere of 4% CO₂ -96% air, 37° C. for 14 days. Medium then wasdecanted and colonies fixed with methanol and stained with 0.1% crystalviolet in 20% methanol. Macroscopic colonies greater than 1 mm indiameter were enumerated. Drug effects were calculated as a percentageof reduction in number of colonies formed in the presence of each drugconcentration compared to the number of colonies formed in the absenceof drugs. Dose-response curves were generated and I₅₀ concentrations forinhibition of plating/colony formation were calculated.

(6) Data Analysis

Dose-response relationships were used to compare drug effects. Thesewere constructed by linearly regressing the percent inhibition ofparameters derived in the preceding sections against log drugconcentrations. The 50 inhibitory (I₅₀) concentrations were calculatedfrom the regression lines using the methods described by Goldstein. SeeGoldstein, A., Biostatistics: An Introductory Text, MacMillan, New York,pp. 156-161 (1964). The three I₅₀ concentrations for inhibition of DNA,RNA and protein synthesis were averaged and were reported in the tables.Along with the I₅₀ concentrations are compared to I₅₀ concentrations forinhibition for HCMV or HSV-1 replication. Compounds for which the ratioof cytotoxicity I₅₀ concentrations to antiviral I₅₀ concentrations (invitro therapeutic index) were greater than 10, were considered forfurther study.

RESULTS (1) Antiviral Activity and Cytotoxicity of Benzimidazoles

Table 1 below summarizes test results from antiviral and cytotoxicityevaluation of the benzimidazoles. These halogen-substituted compoundswere active against HCMV, with a halogen at R₅ being essential forantiviral activity and low cytotoxicity. For example, compounds 45 and52 were active against HCMV in the sub- or low micromolar range and didnot produce cytotoxicity in uninfected cells at concentrations up to 100μM. This potent and selective antiviral activity against HCMV is insharp contrast to the low, apparent activity against this virus of thedichloro compound commonly referred to as DRB which compound wasinitially described by Tamm (I. Tamm, Science 120:847-848, 1954). The R₂and R₃ positions in this compound are substituted by Cl but the R₅position is unsubstituted. Although this compound appears to have weakactivity against HCMV and HSV-1 (Table 1), this activity was observedonly at concentrations which produced cytotoxicity in uninfected cells(Table 1 and FIG. 6). Thus in contrast to compounds disclosed herein,the activity of DRB against HCMV is not actual antiviral activity butrather is a manifestation of cytotoxicity.

Other compounds showing good or better activity than compound 45 againstHCMV and low cytotoxicity are compounds 52, 85, 95, 99 and compound 111(the deoxyribosyl analog of compound 45). Other compounds with activityinclude compounds 61, 81, 83a, and 107. All compounds except 81, 95, and99 also had activity against HSV-1.

                  TABLE 1                                                         ______________________________________                                        50% Inhibitory Concentration (μM)                                          Antiviral Activity Against:                                                                        Cytotoxicity                                             Compound                                                                              HCMV         HSV-1   In Cell Line:                                    Number.sup.a                                                                          Plaque   Yield.sup.b                                                                           Plaque                                                                              HFF.sup.c                                                                           BSC.sup.c                                                                           KB.sup.d                           ______________________________________                                         45      2.8.sup.e                                                                              1.4.sup.e                                                                             151.sup.e                                                                           238.sup.e                                                                          >100   175.sup.e                          52      2.5.sup.e                                                                              0.3.sup.e                                                                             99.sup.e                                                                            100.sup.e  >100                                61     19.sup.e --       27.sup.e                                                                            32.sup.e     9                                 81     11        2.0    >100   32         >100                                83a    30.sup.e 26.sup.e                                                                               21.sup.e                                                                           >100.sup.e   23                                 85      5.7.sup.e                                                                              2.5.sup.e                                                                             55    75.sup.e    109.sup.e                          95      6.0.sup.e                                                                              1.5.sup.e                                                                            >100.sup.e                                                                           100.sup.e   156.sup.e                          99      7.0.sup.e                                                                              2.8.sup.e                                                                            >100.sup.e                                                                           100.sup.e   139.sup.e                         107      1.7.sup.e                                                                              2.0     50    10.sup.e    19                                111     20.sup.e 12       41   >320.sup.e   53                                112     35.sup.e  6       --    32.sup.e    171.sup.e                         DRB.sup.f                                                                             42.sup.e 19.sup.e                                                                               30.sup.e                                                                            24.sup.e    36.sup.e                          ______________________________________                                         .sup.a Number for chemical structure presented in text.                       .sup.b 90% inhibitory concentration (I.sub.90) presented.                     .sup.c Visual cytotoxicity scored on HFF or BSC1 cells at time of HCMV or     HSV1 plaque enumeration.                                                      .sup.d Average percent inhibition of DNA, RNA and protein synthesis or        cell growth determined in KB cells as described in the text.                  .sup.e Average of two to eight replicate experiments.                         .sup.f Initially described by I. Tamm, Science 120:847-848 (1954).       

Table 3 below summarizes test results from antiviral and cytotoxicityevaluation of benzimidazoles and related compounds. These compounds areactive against HCMV and some also are active against HSV-1. For example,compounds 57, 65, 65a, 87, 90, 103, 113, 134, 155, 156, 182, 92, 81c and54 were active against HCMV in the low-micromolar range in either plaqueor yield reduction assays. Although some compounds also showed cytotoxiceffects in some assays, activity against HCMV was separated fromcytotoxicity. All other compounds presented in Table 3 also were activeagainst HCMV but to a lesser extent.

In addition to activity against HCMV, compounds 44, 65a, 113, 134, 182,54, 19, 12c, 13 and 26 also are active against HSV-1.

                  TABLE 3                                                         ______________________________________                                        Antiviral Activity and Cytotoxicity                                           50% Inhibitory Concentration (μM)                                          Antiviral Activity Against:                                                                        Cytotoxicity                                             Compound                                                                              HCMV         HSV-1   In Cell Line:                                    Number.sup.a                                                                          Plaque   Yield.sup.b                                                                           Plaque                                                                              HFF.sup.c                                                                           BSC.sup.c                                                                           KB.sup.d                           ______________________________________                                         7        4                     10                                             12.sup.c                                                                              111.sup.e                                                                             >100.sup.e                                                                             42.sup.e                                                                            77.sup.e    45                                 13      18               79    32                                             19       6.sup.e                                                                                0.01   21.sup.e                                                                            20.sup.e                                       26       2.8      7.0    25    10                                             32       5.1.sup.e      >100.sup.e                                                                           32.sup.e   >100                                41      20                     32                                             41.sup.c                                                                              25                     32                                             44      40               50    32                                             54      22        7.0    25    100   100  >100                                57      30.sup.e                                                                                0.03  >100   44.sup.e    81.sup.e                           65     >100.sup.e                                                                               8.0         >100.sup.e                                      65.sup.a                                                                              32.sup.e                                                                                3.8.sup.e                                                                            149.sup.e                                                                           111.sup.e  >100                                67      127.sup.e                                                                              50.sup.e                                                                             >100  >100.sup.e                                      81.sup.b                                                                              55.sup.e        >100   161.sup.e   100                                81.sup.c                                                                              30        1.0          100                                            87      127.sup.e                                                                              17.sup.e                                                                             >100   161.sup.e                                      90     >100.sup.e                                                                              10.sup.e                                                                             >100   161        >100.sup.e                          92       0.3.sup.e                                                                              0.01         32.sup.e                                      103       0.5.sup.e                                                                              0.4           3.sup.e                                      113      30        0.5    19    32          19                                134      60.sup.e                                                                               19      28   >100.sup.e   58.sup.e                          155      11        0.06  >100   32          70                                156     >100       6.0   >100   100                                           166      69.sup.e                                                                               149.sup.e                                                                            >100   97.sup.e    62                                166a     24              >100  >100  >100  >100                               167      16.sup.e                                                                               168.sup.e                                                                            >100.sup.e                                                                          >100.sup.e                                                                          >100  >100.sup.e                         182      127.sup.e                                                                               1.1.sup.e                                                                            152.sup.e                                                                           99         >142.sup.e                         ______________________________________                                         .sup.a Number for chemical structure presented in text.                       .sup.b 90% inhibitory concentration (I.sub.90) presented.                     .sup.c Visual cytotoxicity scored on HFF or BSC1 cells at time of HCMV or     HSV1 plaque enumeration.                                                      .sup.d Average percent inhibition of DNA, RNA and protein synthesis or        cell growth determined in KB cells as described in the text.                  .sup.e Average of two to eight replicate experiments.                    

(2) Detailed Studies with Compound 45

Because of the potent activity of compound 45 against HCMV and its verylow cytotoxicity, this compound has been studied more extensively. Datain Table 2 below provide evidence that compound 45 is highly specificfor human cytomegalovirus. The data show the compound is highly activeagainst this virus but is less active against herpes simplex type 1,herpes simplex type 2, varicella-zoster virus and mouse cytomegalovirus.

                  TABLE 2                                                         ______________________________________                                        Effect of Compound 45 on the Replication of Herpes Viruses                                       50% Inhibitory                                                                Concentration μM                                        Virus  Cell Line         Virus    Cytotoxicity.sup.a                          ______________________________________                                        HSV-1  Human Foreskin Fibroblasts                                                                       >57     .sup.˜ 140                            HSV-1  Rabbit Kidney     >284     >284                                        HSV-1  Mouse Embryo Fibroblasts                                                                         28      .sup.˜ 140                            HSV-2  Human Foreskin Fibroblasts                                                                       >57     .sup.˜ 140                            HSV-2  Rabbit Kidney     >284     >284                                        HSV-2  Mouse Embryo Fibroblasts                                                                         14      .sup.˜ 140                            VZV    Human Foreskin Fibroblasts                                                                       94      --                                          MCMV   Mouse Embryo Fibroblasts                                                                         57      .sup.˜ 140                            HCMV   Human Foreskin Fibroblasts                                                                        1.4    >>28                                        ______________________________________                                    

Because of its unique potent activity against HCMV, compound 45 wascompared to the known anti-HCMV agent ganciclovir as well as to DRB.FIG. 3 shows that compound 45 is at least as effective as ganciclovir inproducing multiple 10-fold reductions in virus titer. Thus, at aconcentration of 32 μM compound 45 produced a 100,000-fold reduction inthe replication of HCMV. The lack of cytotoxicity of compound 45 in thisantiviral activity range is shown in FIG. 4. Data in this figureestablish that at 32 to 100 μM compound 45 had little or no effect onthe growth of uninfected KB cells. FIG. 5 shows that the inhibitoryeffects produced by a concentration as high as 320 μM could be reversedby simple removal of the drug from uninfected cells. In contrast FIG. 6shows the effects of the known compound DRB were fully inhibitory touninfected cells at 100 μM and these effects could not be reversed byremoval of the drug from culture, thereby establishing the cytoxicity ofthis compound.

The lack of cytotoxicity of compound 45 was further established byplating efficiency experiments. In these experiments, which measure boththe ability of cells to grow and to attach to a substrate, compound 45had no effect at 100 μM.

(3) Additional Studies with Compounds 45 and 52

Because of the potent activity of compounds 45 and 52 against HCMV andtheir very low cytotoxicity, these compounds were studied further.Compound 52 also was compared to the known anti-HCMV agent ganciclovirin yield reduction assays. Compound 52 is more effective thanganciclovir in producing multiple 10-fold reductions in virus titer.Thus, at a concentration of 3.2 μM, compound 52 produced nearly a100,000-fold reduction in the replication of HCMV. In contrast, a 32 μMconcentration of ganciclovir was needed to give a similar reduction inHCMV titer. Separate experiments (Table 1) established a lack ofcytotoxicity of compound 52 in its antiviral activity range. Data inthis figure show that up to 100 μM compound 52 had little or no effecton the growth of uninfected KB cells.

The lack of toxicity of compounds 45 and 52 in their antiviral doserange was further established by examining the effect of both compoundson human bone marrow progenitor cells. Both compounds are at leastsix-fold less toxic to granulocyte/macrophage progenitor cells than isganciclovir. Compound 45 is at least four-fold less toxic to erythroidprogenitor cells than is ganciclovir, compound 52 is nearly three-foldless toxic. Thus, both compounds are more active against HCMV than isganciclovir and both are less toxic to human cells.

(4) Drug Combination and Synergy

Compounds of the invention, such as compounds 45 and 52, provideadditional advantages when used in combination with other antiviraldrugs. For example, combination of compound 45 with the known antiviraldrug ganciclovir results in greater activity (i.e. synergy) againsthuman cytomegalovirus (HCMV) than the use of either agent alone whenused at concentrations (approximately 0.1 to 10 μM) most likely achievedin vivo by the administration of therapeutic amounts of each drug. Athigher concentrations (approximately 3 to 10 μM) compound 45 also issynergistic against HCMV when used with another known antiviral drug,acyclovir. Likewise the combination of compound 52 with ganciclovir oracyclovir results in synergistic activity against HCMV.

Compounds of the invention, such as 45 and 52, could thus be used totreat HCMV infections in AIDS patients already receiving the antiviraldrug zidovudine (AZT). Combination of either compound 45 or 52 with AZTprovides the advantage of less toxicity over the combination ofganciclovir with AZT. The combination of compound 45 or 52 with AZTproduces less cytotoxicity (i.e. antagonism) in cultured human cellsthan either agent used alone. In contrast, combination of ganciclovirwith AZT produces greater cytotoxicity in human cells than the use ofeither of these drugs alone.

(5) Activity of Ester Prodrugs

Prodrugs of polysubstituted benzimidazoles can be useful for oraladministration. For example, ester prodrugs of compounds 45 (compounds42 and 42a) and 52 (compounds 52a and 52b), respectively, have beenprepared and are active against HCMV.

The foregoing discussion discloses and describes merely exemplaryembodiments of the present invention. One skilled in the art willreadily recognize from such discussion, and from the accompanyingdrawings and claims, that various changes, modifications and variationscan be made therein without departing from the spirit and scope of theinvention as defined in the following claims. Any discrepancy incompound nomenclature in the specification and claims herein should beresolved in deference to the structures and substituents set forth inthe charts of FIGS. 1 and 2.

What is claimed is:
 1. An antiviral compound selected from the groupconsisting of compounds having the following formula, andpharmaceutically acceptable salts thereof: ##STR6## wherein: R₁ is H, R₂is Cl, R₃ is Cl, R₄ is H, R₅ is NH₂ and R₆ is2'-deoxy-β-D-erythropentofuranosyl (denoted compound 113 in the text);R₁is H, R₂ is Cl, R₃ is Cl, R₄ is H, R₅ is Cl and R₆ isβ-D-arabinofuranosyl (denoted compound 134 in the text); and R₁ is H, R₂is Cl, R₃ is Cl, R₄ is H, R₅ is Cl and R₆ is5'-O-acetyl-β-D-ribofuranosyl (denoted compound 42a in the text).
 2. Theantiviral compound of claim 1, wherein R₁ is H, R₂ is Cl, R₃ is Cl, R₄is H, R₅ is NH₂ and R₆ is 2'-deoxy-β-D-erythro-pentofuranosyl (denotedcompound 113 in the text).
 3. The antiviral compound of claim 1, whereinR₁ is H, R₂ is Cl, R₃ is Cl, R₄ is H, R₅ is Cl and R₆ isβ-D-arabinofuranosyl (denoted compound 134 in the text).
 4. Theantiviral compound of claim 1, wherein R₁ is H, R₂ is Cl, R₃ is Cl, R₄is H, R₅ is Cl and R₆ is 5'-O-acetyl-β-D-ribofuranosyl (denoted compound42a in the text).
 5. An antiviral composition comprising apharmaceutically acceptable carrier and an compound selected from thegroup consisting of compounds having the following formula, andpharmaceutically acceptable salts thereof: ##STR7## wherein: R₁ is H, R₂is Cl, R₃ is Cl, R₄ is H, R₅ is NH₂ and R₆ is2'-deoxy-β-D-erythropentofuranosyl (denoted compound 113 in the text);R₁is H, R₂ is Cl, R₃ is Cl, R₄ is H, R₅ is Cl and R₆ isβ-D-arabinofuranosyl (denoted compound 134 in the text); R₁ is H, R₂ isCl, R₃ is Cl, R₄ is H, R₅ is Cl and R₆ is 5'-O-acetyl-β-D-ribofuranosyl(denoted compound 42a in the text).
 6. The antiviral composition ofclaim 5, wherein R₁ is H, R₂ is Cl, R₃ is Cl, R₄ is H, R₅ is NH₂ and R₆is 2'-deoxy-β-D-erythro-pentofuranosyl (denoted compound 113 in thetext).
 7. The antiviral composition of claim 5, wherein R₁ is H, R₂ isCl, R₃ is Cl, R₄ is H, R₅ is Cl and R₆ is β-D-arabinofuranosyl (denotedcompound 134 in the text).
 8. The antiviral composition of claim 5,wherein R₁ is H, R₂ is Cl, R₃ is Cl, R₄ is H, R₅ is Cl and R₆ is5'-O-acetyl-β-D-ribofuranosyl (denoted compound 42a in the text).
 9. Theantiviral composition of claim 5, further comprising ganciclovir.
 10. Amethod for treating a herpes viral infection comprising administering tothe infected host a therapeutically effective amount of a compound, or apharmaceutically acceptable salt or formulation thereof, selected fromthe group consisting of compounds having the following formula: ##STR8##wherein: R₁ is H, R₂ is Cl, R₃ is Cl, R₄ is H, R₅ is NH₂ and R₆ is2'-deoxy-β-D-erythropentofuranosyl (denoted compound 113 in the text);R₁is H, R₂ is Cl, R₃ is Cl, R₄ is H, R₅ is Cl and R₆ isβ-D-arabinofuranosyl (denoted compound 134 in the text); R₁ is H, R₂ isCl, R₃ is Cl, R₄ is H, R₅ is Cl and R₆ is 5'-O-acetyl-β-D-ribofuranosyl(denoted compound 42a in the text); and R₁ is H, R₂ is Cl, R₃ is Cl, R₄is H, R₅ is Cl and R₆ is 2',3',5'-tri-O-acetyl-β-D-ribofuranosyl(denoted compound 42 in the text).
 11. The method of claim 10, whereinR₁ is H, R₂ is Cl, R₃ is Cl, R₄ is H, R₅ is NH₂ and R₆ is2'-deoxy-β-D-erythro-pentofuranosyl (denoted compound 113 in the text).12. The method of claim 10, wherein R₁ is H, R₂ is Cl, R₃ is Cl, R₄ isH, R₅ is Cl and R₆ is β-D-arabinofuranosyl (denoted compound 134 in thetext).
 13. The method of claim 10, wherein R₁ is H, R₂ is Cl, R₃ is Cl,R₄ is H, R₅ is Cl and R₆ is 5'-O-acetyl-β-D-ribofuranosyl (denotedcompound 42a in the text).
 14. The method of claim 10, wherein R₁ is H,R₂ is Cl, R₃ is Cl, R₄ is H, R₅ is Cl and R₆ is2',3',5'-tri-O-acetyl-β-D-ribofuranosyl (denoted compound 42 in thetext).